Nucleic acid molecules encoding chimeric antigen receptors targeting b-cell maturation antigen and uses thereof

ABSTRACT

The presently disclosed subject matter provides for methods and compositions for treating multiple myeloma. It relates to chimeric antigen receptors (CARs) that specifically target B cell maturation antigen (BMCA), and immunoresponsive cells comprising such CARs. The presently disclosed BMCA-specific CARs have enhanced immune-activating properties, including anti-tumor activity.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Divisional of U.S. patent application Ser. No.15/613,638, filed Jun. 5, 2017, which is a Continuation of InternationalApplication Serial No. PCT/US2015/064112, filed Dec. 4, 2015, whichclaims priority to U.S. Provisional Patent Application Ser. No.62/088,309, filed Dec. 5, 2014, the contents of each of which areincorporated by reference in their entirety, and to each of whichpriority is claimed.

SEQUENCE LISTING

The specification further incorporates by reference the Sequence Listingsubmitted herewith via EFS on Apr. 9, 2020. Pursuant to 37 C.F.R. §1.52(e)(5), the Sequence Listing text file, identified as0727341034_SL.txt, is 234,902 bytes and was created on Apr. 9, 2020. TheSequence Listing electronically filed herewith, does not extend beyondthe scope of the specification and thus does not contain new matter.

INTRODUCTION

The presently disclosed subject matter provides for methods andcompositions for treating cancer. It relates to chimeric antigenreceptors (CARs) that specifically target B-cell maturation antigen(BCMA), immunoresponsive cells comprising such CARs, and methods ofusing such cells for treating cancer (e.g., multiple myeloma).

BACKGROUND OF THE INVENTION

Cell-based immunotherapy is a therapy with curative potential for thetreatment of cancer. T cells and other immune cells may be modified totarget tumor antigens through the introduction of genetic materialcoding for artificial or synthetic receptors for antigen, termedChimeric Antigen Receptors (CARs), specific to selected antigens.Targeted T cell therapy using CARs has shown recent clinical success intreating hematologic malignancies.

Multiple myeloma (MM) is the second most common hematologic malignancy.⁸Approximately 25% of patients have high-risk cytogenetics, whichportends a median survival of less then 2 years.^(9,10) While recentstrides have been made, regardless of cytogenetics, the disease is stillconsidered incurable outside the immuno-therapeutic graft versus myeloma(GvM) effect of an allogeneic transplant. However, allogeneictransplants are limited by ineligibility and high rates oftransplant-associated morbidity and mortality.¹¹ Similar to the GvMeffect, a potentially curative T cell effect may be achieved withminimal toxicity through autologous adoptive T cell therapy.

Myeloma may be an ideal disease to test adoptive T cell therapy. First,as indicated above, allogeneic transplants demonstrate that the T cellcan be a curative treatment, even with minimal or no concomitantchemotherapy such as after non-myeloablative transplants orpost-transplantation donor lymphocyte infusions. Second, conditioningchemotherapy, possibly through the mechanism of depleting regulatory Tcells (Tregs), enhances the efficacy of adoptive T cell therapy,^(4,12)as such, the immediate post-autologous transplant period could be anoptimal time to administer T cells, and myeloma is one of the fewdiseases where autologous stem cell transplantation is the standard ofcare. Third, the immunomodulatory drug lenalidomide may improve CARbased therapy, as has been shown in mice,¹³ and lenalidomide is commonlyused to treat MM. Fourth, adoptive T cell therapy works best in bonemarrow predominant disease such as ALL,^(6,7) when compared to solidtumors or extra-medullary CLL,⁴ and similar to ALL, myeloma is a diseaseof the bone marrow.

While there are various reasons to expect that adoptive T cell therapymay work well in MM, expanding adoptive T cell therapy to myeloma alsoposes unique challenges. Unlike other B-cell malignancies, CD19expression is seen in only 2% of myeloma patients.¹⁴ Furthermore, unlikeCD19, the common extracellular immunophenotypic markers in myeloma(CD138, CD38, and CD56) are all co-expressed on other essential celltypes, and we predict CARs to any of these targets would lead tounacceptable “off tumor, on target” toxicity⁶ which can be fatal even intargets where antibodies are well tolerated, as was the case with a HER2targeted CAR.¹⁵ Accordingly, there are needs for novel therapeuticstrategies to design CARs targeting antigens that are highly expressedin MM cells and limited expression in normal tissues for treatingmultiple myeloma, which strategies capable of inducing potent tumoreradication with minimal toxicity and immunogenicity.

SUMMARY OF THE INVENTION

The presently disclosed subject matter generally provides chimericantigen receptors (CARs) that specifically target B-cell maturationantigen (BCMA), immunoresponsive cells comprising such CARs, and uses ofthese CARs and immunoresponsive cells for treating multiple myeloma.

The presently disclosed subject matter provides CARs. In onenon-limiting example, the CAR comprises an extracellular antigen-bindingdomain, a transmembrane domain and an intracellular domain, where theextracellular antigen-binding domain is a human single-chain variablefragment (scFv) that specifically binds to B cell maturation antigen(BMCA). In certain embodiments, the human scFv specifically binds toBCMA with a binding affinity (K_(D)) of from about 1×10⁻⁹M to about3×10⁻⁶ M. In certain embodiments, the human scFv specifically binds toBCMA with a binding affinity (K_(D)) of from about 1×10⁻⁹M to about1×10⁻⁸M.

In certain embodiments, the human scFv comprises a heavy chain variableregion comprising an amino acid sequence that is at least about 80%,about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%,about 94%, about 95%, about 96%, about 97%, about 98% or about 99%homologous to an amino acid sequence selected from the group consistingof SEQ ID NOS: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57,61, and 65.

In certain embodiments, the human scFv comprises a light chain variableregion comprising an amino acid sequence that is at least about 80%,about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%,about 94%, about 95%, about 96%, about 97%, about 98% or about 99%homologous to an amino acid sequence selected from the group consistingof SEQ ID NOS: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58,62, and 66.

In certain embodiments, the human scFv comprises (a) a heavy chainvariable region comprising an amino acid sequence that is at least about80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%,about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99%homologous to an amino acid sequence selected from the group consistingof SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57,61, and 65; and (b) a light chain variable region comprising an aminoacid sequence that is at least about 80%, about 81%, about 82%, about83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%,about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about96%, about 97%, about 98% or about 99% homologous to an amino acidsequence selected from the group consisting of SEQ ID NOS: 2, 6, 10, 14,18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, and 66.

In certain embodiments, the human scFv comprises a heavy chain variableregion comprising an amino acid sequence selected from the groupconsisting of SEQ ID NOS: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45,49, 53, 57, 61, and 65, and conservative modifications thereof.

In certain embodiments, the human scFv comprises a light chain variableregion comprising an amino acid sequence selected from the groupconsisting of SEQ ID NOS: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46,50, 54, 58, 62, and 66, and conservative modifications thereof.

In certain embodiments, the human scFv comprises (a) a heavy chainvariable region comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOS: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41,45, 49, 53, 57, 61, and 65, and conservative modifications thereof, and(b) a light chain variable region comprising an amino acid sequenceselected from the group consisting of SEQ ID NOS: 2, 6, 10, 14, 18, 22,26, 30, 34, 38, 42, 46, 50, 54, 58, 62, and 66, and conservativemodifications thereof.

In certain embodiments, the human scFv comprises a heavy chain variableregion comprising amino acids having a sequence selected from the groupconsisting of: SEQ ID NOS:1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45,49, 53, 57, 61, and 65. In certain embodiments, the human scFv comprisesa light chain variable region comprising amino acids having a sequenceselected from the group consisting of: SEQ ID NOS:2, 6, 10, 14, 18, 22,26, 30, 34, 38, 42, 46, 50, 54, 58, 62, and 66. In certain embodiments,the human scFv comprises a heavy chain variable region comprising aminoacids having the sequence set forth in SEQ ID NO:21. In certainembodiments, the human scFv comprises a heavy chain variable regioncomprising amino acids having the sequence set forth in SEQ ID NO:53. Incertain embodiments, the human scFv comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:57. In certain embodiments, the human scFv comprises a heavy chainvariable region comprising amino acids having the sequence set forth inSEQ ID NO:61. In certain embodiments, the human scFv comprises a heavychain variable region comprising amino acids having the sequence setforth in SEQ ID NO:65. In certain embodiments, the human scFv comprisesa light chain variable region comprising amino acids having the sequenceset forth in SEQ ID NO:22. In certain embodiments, the human scFvcomprises a light chain variable region comprising amino acids havingthe sequence set forth in SEQ ID NO:54. In certain embodiments, thehuman scFv comprises a light chain variable region comprising aminoacids having the sequence set forth in SEQ ID NO:58. In certainembodiments, the human scFv comprises a light chain variable regioncomprising amino acids having the sequence set forth in SEQ ID NO:62. Incertain embodiments, the human scFv comprises a light chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:66. In certain embodiments, the extracellular antigen-binding domaincomprises (a) a heavy chain variable region comprising amino acidshaving a sequence set forth in SEQ ID NO:1, and a light chain variableregion comprising amino acids having a sequence set forth in SEQ IDNO:2; (b) a heavy chain variable region comprising amino acids having asequence set forth in SEQ ID NO:5, and a light chain variable regioncomprising amino acids having a sequence set forth in SEQ ID NO:6; (c) aheavy chain variable region comprising amino acids having a sequence setforth in SEQ ID NO:9, and a light chain variable region comprising aminoacids having a sequence set forth in SEQ ID NO:10; (d) a heavy chainvariable region comprising amino acids having a sequence set forth inSEQ ID NO:13, and a light chain variable region comprising amino acidshaving a sequence set forth in SEQ ID NO:14; (e) a heavy chain variableregion comprising amino acids having a sequence set forth in SEQ IDNO:17, and a light chain variable region comprising amino acids having asequence set forth in SEQ ID NO:18; (f) a heavy chain variable regioncomprising amino acids having a sequence set forth in SEQ ID NO:21, anda light chain variable region comprising amino acids having a sequenceset forth in SEQ ID NO:22; (g) a heavy chain variable region comprisingamino acids having a sequence set forth in SEQ ID NO:25, and a lightchain variable region comprising amino acids having a sequence set forthin SEQ ID NO:26; (h) a heavy chain variable region comprising aminoacids having a sequence set forth in SEQ ID NO:29, and a light chainvariable region comprising amino acids having a sequence set forth inSEQ ID NO:30; (i) a heavy chain variable region comprising amino acidshaving a sequence set forth in SEQ ID NO:33, and a light chain variableregion comprising amino acids having a sequence set forth in SEQ IDNO:34; (j) a heavy chain variable region comprising amino acids having asequence set forth in SEQ ID NO:37, and a light chain variable regioncomprising amino acids having a sequence set forth in SEQ ID NO:38; (k)a heavy chain variable region comprising amino acids having a sequenceset forth in SEQ ID NO:41, and a light chain variable region comprisingamino acids having a sequence set forth in SEQ ID NO:42; (l) a heavychain variable region comprising amino acids having a sequence set forthin SEQ ID NO:45, and a light chain variable region comprising aminoacids having a sequence set forth in SEQ ID NO:46; (m) a heavy chainvariable region comprising amino acids having a sequence set forth inSEQ ID NO:49, and a light chain variable region comprising amino acidshaving a sequence set forth in SEQ ID NO:50; (n) a heavy chain variableregion comprising amino acids having a sequence set forth in SEQ IDNO:53, and a light chain variable region comprising amino acids having asequence set forth in SEQ ID NO:54; (o) a heavy chain variable regioncomprising amino acids having a sequence set forth in SEQ ID NO:57, anda light chain variable region comprising amino acids having a sequenceset forth in SEQ ID NO:58; (p) a heavy chain variable region comprisingamino acids having a sequence set forth in SEQ ID NO:61, and a lightchain variable region comprising amino acids having a sequence set forthin SEQ ID NO:62; or (q) a heavy chain variable region comprising aminoacids having a sequence set forth in SEQ ID NO:65, and a light chainvariable region comprising amino acids having a sequence set forth inSEQ ID NO:66. In certain embodiments, the human scFv comprises a heavychain variable region comprising amino acids having the sequence setforth in SEQ ID NO:21; and a light chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:22. In certainembodiments, the human scFv comprises a heavy chain variable regioncomprising amino acids having the sequence set forth in SEQ ID NO:53;and a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:54. In certain embodiments, the humanscFv comprises a heavy chain variable region comprising amino acidshaving the sequence set forth in SEQ ID NO:57; and a light chainvariable region comprising amino acids having the sequence set forth inSEQ ID NO:58. In certain embodiments, the human scFv comprises a heavychain variable region comprising amino acids having the sequence setforth in SEQ ID NO:61; and a light chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:62. In certainembodiments, the human scFv comprises a heavy chain variable regioncomprising amino acids having the sequence set forth in SEQ ID NO:65;and a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:66.

In certain non-limiting embodiments, the human scFv comprises both ofsaid heavy and light chains, optionally with a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. For example, in certain non-limitingembodiments, the human scFv comprises (i) a heavy chain variable regioncomprising amino acids having the sequence set forth in SEQ ID NO:53 and(ii) a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:54, optionally with (iii) a linkersequence, for example a linker peptide, between the heavy chain variableregion and the light chain variable region. In certain embodiments, thehuman scFv comprises (i) a heavy chain variable region comprising aminoacids having the sequence set forth in SEQ ID NO:21 and (ii) a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:22, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the human scFvcomprises (i) a heavy chain variable region comprising amino acidshaving the sequence set forth in SEQ ID NO:57 and (ii) a light chainvariable region comprising amino acids having the sequence set forth inSEQ ID NO:58, optionally with (iii) a linker sequence, for example alinker peptide, between the heavy chain variable region and the lightchain variable region. In certain embodiments, the human scFv comprises(i) a heavy chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:61 and (ii) a light chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:62, optionally with (iii) a linker sequence, for example a linkerpeptide, between the heavy chain variable region and the light chainvariable region. In certain embodiments, the human scFv comprises (i) aheavy chain variable region comprising amino acids having the sequenceset forth in SEQ ID NO:65 and (ii) a light chain variable regioncomprising amino acids having the sequence set forth in SEQ ID NO:66,optionally with (iii) a linker sequence, for example a linker peptide,between the heavy chain variable region and the light chain variableregion.

In certain embodiments, the human scFv comprises (a) a heavy chainvariable region CDR3 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOS: 91, 97, 103, 109, 115, 121, 127, 133,139, 145, 151, 157, 163, 169, 175, 181, and 187; and (b) a light chainvariable region CDR3 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOS: 94, 100, 106, 112, 118, 124, 130, 136,142, 148, 154, 160, 166, 172, 178, 184, and 190.

In certain embodiments, the human scFv comprises (a) a heavy chainvariable region CDR2 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 90, 96, 102, 108, 114, 120, 126, 132,138, 144, 150, 156, 162, 168, 174, 180, and 186, and conservativemodifications thereof; and (b) a light chain variable region CDR2comprising an amino acid sequence selected from the group consisting ofSEQ ID NOS: 93, 99, 105, 111, 117, 123, 129, 135, 141, 147, 153, 159,165, 171, 177, 183, and 189, and conservative modifications thereof.

In certain embodiments, the human scFv comprises (a) a heavy chainvariable region CDR1 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 89, 95, 101, 107, 113, 119, 125, 131,137, 143, 149, 155, 161, 167, 173, 179, and 185, and conservativemodifications thereof; and (b) a light chain variable region CDR1comprising an amino acid sequence selected from the group consisting ofSEQ ID NOS: 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158,164, 170, 176, 182, and 188, and conservative modifications thereof. Incertain embodiments, the human scFv comprises: (a) a heavy chainvariable region CDR1 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOS: 89, 95, 101, 107, 113, 119, 125, 131,137, 143, 149, 155, 161, 167, 173, 179, and 185; (b) a heavy chainvariable region CDR2 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 90, 96, 102, 108, 114, 120, 126, 132,138, 144, 150, 156, 162, 168, 174, 180, and 186; (c) a heavy chainvariable region CDR3 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOS: 91, 97, 103, 109, 115, 121, 127, 133,139, 145, 151, 157, 163, 169, 175, 181, and 187; (d) a light chainvariable region CDR1 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 92, 98, 104, 110, 116, 122, 128, 134,140, 146, 152, 158, 164, 170, 176, 182, and 188; (e) a light chainvariable region CDR2 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 93, 99, 105, 111, 117, 123, 129, 135,141, 147, 153, 159, 165, 171, 177, 183, and 189; and (f) a light chainvariable region CDR3 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 94, 100, 106, 112, 118, 124, 130, 136,142, 148, 154, 160, 166, 172, 178, 184, and 190. In certain embodiments,the human scFv comprises (a) a heavy chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 89 orconservative modifications thereof; a heavy chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 90 orconservative modifications thereof; and a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:91 or conservative modifications thereof; (b) a heavy chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 95 or conservative modifications thereof; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 96 or conservative modifications thereof; and a heavy chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 97 or conservative modifications thereof; (c) aheavy chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 101 or conservative modificationsthereof; a heavy chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 102 or conservativemodifications thereof; and a heavy chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 103 orconservative modifications thereof; (d) a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:107 or conservative modifications thereof; a heavy chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:108 or conservative modifications thereof; and a heavy chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 109 or conservative modifications thereof; (e) a heavy chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 113 or conservative modifications thereof; a heavychain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO: 114 or conservative modifications thereof; and aheavy chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 115 or conservative modificationsthereof; (f) a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 119 or conservativemodifications thereof; a heavy chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO: 120 orconservative modifications thereof; and a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:121 or conservative modifications thereof; (g) a heavy chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 125 or conservative modifications thereof; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 126 or conservative modifications thereof; and a heavy chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 127 or conservative modifications thereof; (h) aheavy chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 131 or conservative modificationsthereof; a heavy chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 132 or conservativemodifications thereof; and a heavy chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 133 orconservative modifications thereof; (i) a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:137 or conservative modifications thereof; a heavy chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:138 or conservative modifications thereof; and a heavy chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 139 or conservative modifications thereof; (j) a heavy chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 143 or conservative modifications thereof; a heavychain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO: 144 or conservative modifications thereof; and aheavy chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 145 or conservative modificationsthereof; (k) a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 149 or conservativemodifications thereof; a heavy chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO: 150 orconservative modifications thereof; and a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:151 or conservative modifications thereof; (l) a heavy chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 155 or conservative modifications thereof; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 156 or conservative modifications thereof; and a heavy chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 157 or conservative modifications thereof; (m) aheavy chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 161 or conservative modificationsthereof; a heavy chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 162 or conservativemodifications thereof; and a heavy chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 163 orconservative modifications thereof; (n) a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:167 or conservative modifications thereof; a heavy chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:168 or conservative modifications thereof; and a heavy chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 169 or conservative modifications thereof; (o) a heavy chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 173 or conservative modifications thereof; a heavychain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO: 174 or conservative modifications thereof; and aheavy chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 175 or conservative modificationsthereof; (p) a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 179 or conservativemodifications thereof; a heavy chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:180 orconservative modifications thereof; and a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:181 or conservative modifications thereof; or (q) a heavy chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 185 or conservative modifications thereof; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 186 or conservative modifications thereof; and a heavy chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 187 or conservative modifications thereof. Incertain embodiments, the human scFv comprises: a heavy chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 167; a heavy chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 168; and a heavy chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 169. In certain embodiments, the human scFvcomprises: a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 119; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 120; and a heavy chain variable region CDR3 comprising aminoacids having the sequence set forth in SEQ ID NO: 121. In certainembodiments, the human scFv comprises: a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:173; a heavy chain variable region CDR2 comprising amino acids havingthe sequence set forth in SEQ ID NO: 174; and a heavy chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 175. In certain embodiments, the human scFv comprises: a heavychain variable region CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 179; a heavy chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 180;and a heavy chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 181. In certain embodiments, the humanscFv comprises: a heavy chain variable region CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 185; a heavy chainvariable region CDR2 comprising amino acids having the sequence setforth in SEQ ID NO: 186; and a heavy chain variable region CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 187.

In certain embodiments, the human scFv comprises (a) a light chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 92 or conservative modifications thereof; a lightchain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO:93 or conservative modifications thereof; and alight chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 94 or conservative modificationsthereof; (b) a light chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 98 or conservativemodifications thereof; a light chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:99 orconservative modifications thereof; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:100 or conservative modifications thereof; (c) a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 104 or conservative modifications thereof; a light chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO:105 or conservative modifications thereof; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 106 or conservative modifications thereof; (d) alight chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 110 or conservative modificationsthereof; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO:111 or conservativemodifications thereof; and a light chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 112 orconservative modifications thereof; (e) a light chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:116 or conservative modifications thereof; a light chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ IDNO:117 or conservative modifications thereof; and a light chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 118 or conservative modifications thereof; (f) a light chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 122 or conservative modifications thereof; a lightchain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO:123 or conservative modifications thereof; and alight chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 124 or conservative modificationsthereof; (g) a light chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 128 or conservativemodifications thereof; a light chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:129 orconservative modifications thereof; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:130 or conservative modifications thereof; (h) a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 134 or conservative modifications thereof; a light chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO:135 or conservative modifications thereof; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 136 or conservative modifications thereof; (i) alight chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 140 or conservative modificationsthereof; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO:141 or conservativemodifications thereof; and a light chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 142 orconservative modifications thereof; (j) a light chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:146 or conservative modifications thereof; a light chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ IDNO:147 or conservative modifications thereof; and a light chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 148 or conservative modifications thereof; (k) a light chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 152 or conservative modifications thereof; a lightchain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO:153 or conservative modifications thereof; and alight chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 154 or conservative modificationsthereof; (l) a light chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 158 or conservativemodifications thereof; a light chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:159 orconservative modifications thereof; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:160 or conservative modifications thereof; (m) a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 164 or conservative modifications thereof; a light chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO:165 or conservative modifications thereof; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 166 or conservative modifications thereof; (n) alight chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 170 or conservative modificationsthereof; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 171 or conservativemodifications thereof; and a light chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 172 orconservative modifications thereof; (o) a light chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:176 or conservative modifications thereof; a light chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:177 or conservative modifications thereof; and a light chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 178 or conservative modifications thereof; (p) a light chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 182 or conservative modifications thereof; a lightchain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO: 183 or conservative modifications thereof; and alight chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 184 or conservative modificationsthereof; or (q) a light chain variable region CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 188 or conservativemodifications thereof; a light chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO: 189 orconservative modifications thereof; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:190 or conservative modifications thereof. In certain embodiments, thehuman scFv comprises: a light chain variable region CDR1 comprisingamino acids having the sequence set forth in SEQ ID NO: 122; a lightchain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO: 123; and a light chain variable region CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 124.In certain embodiments, the human scFv comprises: a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 170; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 171; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 172. In certain embodiments, the human scFvcomprises: a light chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 176; a light chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 177; and a light chain variable region CDR3 comprising aminoacids having the sequence set forth in SEQ ID NO: 178. In certainembodiments, the human scFv comprises: a light chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:182; a light chain variable region CDR2 comprising amino acids havingthe sequence set forth in SEQ ID NO: 183; and a light chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 184. In certain embodiments, the human scFv comprises: a lightchain variable region CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 188; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 189;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 190.

In certain embodiments, the human scFv comprises: (a) a heavy chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 89; a heavy chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO: 90; a heavychain variable region CDR3 comprising amino acids having the sequenceset forth in SEQ ID NO: 91; a light chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 92; alight chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 93; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:94; (b) a heavy chain variable region CDR1 comprising amino acids havingthe sequence set forth in SEQ ID NO: 95; a heavy chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:96; a heavy chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 97; a light chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 98; alight chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 99; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:100; (c) a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 101; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 102; a heavy chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 103; a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 104; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 105; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 106; (d) a heavy chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 107;a heavy chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 108; a heavy chain variable region CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 109;a light chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 110; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 111;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 112; (e) a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:113; a heavy chain variable region CDR2 comprising amino acids havingthe sequence set forth in SEQ ID NO: 114; a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:115; a light chain variable region CDR1 comprising amino acids havingthe sequence set forth in SEQ ID NO: 116; a light chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:117; and a light chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 118; (f) a heavy chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 119; a heavy chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO: 120; a heavychain variable region CDR3 comprising amino acids having the sequenceset forth in SEQ ID NO: 121; a light chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 122;a light chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 123; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:124; (g) a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 125; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 126; a heavy chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 127; a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 128; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 129; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 130; (h) a heavy chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 131;a heavy chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 132; a heavy chain variable region CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 133;a light chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 134; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 135;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 136; (i) a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:137; a heavy chain variable region CDR2 comprising amino acids havingthe sequence set forth in SEQ ID NO: 138; a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:139; a light chain variable region CDR1 comprising amino acids havingthe sequence set forth in SEQ ID NO: 140; a light chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:141; and a light chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 142; (j) a heavy chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 143; a heavy chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO: 144; a heavychain variable region CDR3 comprising amino acids having the sequenceset forth in SEQ ID NO: 145; a light chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 146;a light chain variable region CDR2 comprising amino acids having thesequence set forth in SE ID NO: 147; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:148; (k) a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 149; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 150; a heavy chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 151; a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 152; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 153; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 154; (l) a heavy chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 155;a heavy chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 156; a heavy chain variable region CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 157;a light chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 158; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 159;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 160; (m) a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:161; a heavy chain variable region CDR2 comprising amino acids havingthe sequence set forth in SEQ ID NO: 162; a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:163; a light chain variable region CDR1 comprising amino acids havingthe sequence set forth in SEQ ID NO: 164; a light chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ IDNO:165; and a light chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 166; (n) a heavy chainvariable region CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 167; a heavy chain variable region CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO: 168; a heavychain variable region CDR3 comprising amino acids having the sequenceset forth in SEQ ID NO: 169; a light chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 170;a light chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 171; and a light chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:172; (o) a heavy chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 173; a heavy chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 174; a heavy chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 175; a light chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 176; a light chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 177; and a light chainvariable region CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 178; (p) a heavy chain variable region CDR1comprising amino acids having the sequence set forth in SEQ ID NO: 179;a heavy chain variable region CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 180; a heavy chain variable region CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 181;a light chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 182; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 183;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 184; or (q) a heavy chain variableregion CDR1 comprising amino acids having the sequence set forth in SEQID NO: 185; a heavy chain variable region CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO: 186; a heavy chain variableregion CDR3 comprising amino acids having the sequence set forth in SEQID NO: 187; a light chain variable region CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO: 188; a light chain variableregion CDR2 comprising amino acids having the sequence set forth in SEQID NO: 189; and a light chain variable region CDR3 comprising aminoacids having the sequence set forth in SEQ ID NO: 190. In certainembodiments, the human scFv comprises: a heavy chain variable regionCDR1 comprising amino acids having the sequence set forth in SEQ ID NO:119; a heavy chain variable region CDR2 comprising amino acids havingthe sequence set forth in SEQ ID NO: 120; a heavy chain variable regionCDR3 comprising amino acids having the sequence set forth in SEQ ID NO:121; a light chain variable region CDR1 comprising amino acids havingthe sequence set forth in SEQ ID NO: 122; a light chain variable regionCDR2 comprising amino acids having the sequence set forth in SEQ ID NO:123; and a light chain variable region CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO: 124. In certain embodiments,the human scFv comprises: a heavy chain variable region CDR1 comprisingamino acids having the sequence set forth in SEQ ID NO: 167; a heavychain variable region CDR2 comprising amino acids having the sequenceset forth in SEQ ID NO: 168; a heavy chain variable region CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 169;a light chain variable region CDR1 comprising amino acids having thesequence set forth in SEQ ID NO: 170; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 171;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 172. In certain embodiments, the humanscFv comprises: a heavy chain variable region CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 173; a heavy chainvariable region CDR2 comprising amino acids having the sequence setforth in SEQ ID NO: 174; a heavy chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 175; a lightchain variable region CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 176; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 177;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 178. In certain embodiments, the humanscFv comprises: a heavy chain variable region CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 179; a heavy chainvariable region CDR2 comprising amino acids having the sequence setforth in SEQ ID NO: 180; a heavy chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 181; a lightchain variable region CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 182; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 183;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 184. In certain embodiments, the humanscFv comprises: a heavy chain variable region CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 185; a heavy chainvariable region CDR2 comprising amino acids having the sequence setforth in SEQ ID NO: 186; a heavy chain variable region CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 187; a lightchain variable region CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 188; a light chain variable region CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 189;and a light chain variable region CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 190.

In certain embodiments, the BCMA comprises the amino acid sequence setforth in SEQ ID NO:71. In certain embodiments, the human scFv binds toan epitope region comprising amino acids 14-22 of SEQ ID NO:71. Incertain embodiments, the human scFv that binds to an epitope regioncomprising amino acids 14-22 of SEQ ID NO:71 comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:21 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:22, In certain embodiments, the human scFv that binds to an epitoperegion comprising amino acids 14-22 of SEQ ID NO:71 comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:119, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:120, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:121, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:122, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:123, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:124.

In accordance with the presently disclosed subject matter, theextracellular antigen-binding domain is covalently joined to atransmembrane domain. The extracellular antigen-binding domain cancomprise a signal peptide that is covalently joined to the 5′ terminusof the extracellular antigen-binding domain. In certain embodiments, thetransmembrane domain of the CAR comprises a CD8 polypeptide, a CD28polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide,an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide,a synthetic peptide (not based on a protein associated with the immuneresponse), or a combination thereof. In certain embodiments, thetransmembrane domain comprises a CD8 polypeptide. In certainembodiments, the transmembrane domain comprises a CD28 polypeptide.

In accordance with the presently disclosed subject matter, theintracellular domain comprises a CD3ζ polypeptide. In certainembodiments, the intracellular domain further comprises at least onesignaling region. In certain embodiments, the at least one signalingregion comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, a PD-1polypeptide, a CTLA-4 polypeptide, a LAG-3 polypeptide, a 2B4polypeptide, a BTLA polypeptide, a synthetic peptide (not based on aprotein associated with the immune response), or a combination thereof.In certain embodiments, the signaling region is a co-stimulatorysignaling region. In certain embodiments, the co-stimulatory signalingregion comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combinationthereof. In certain embodiments, the at least one co-stimulatorysignaling region comprises a CD28 polypeptide. In certain embodiments,the at least one signaling region comprises a 4-1BB polypeptide. In onespecific non-limiting embodiment, the transmembrane domain comprises aCD28 polypeptide, the intracellular domain comprises a CD3ζ polypeptide,and the at least one signaling domain comprises a CD28 polypeptide.

In certain embodiments, the CAR is recombinantly expressed. The CAR canbe expressed from a vector. In certain embodiments, the vector is aγ-retroviral rector.

The presently disclosed subject matter also provides isolatedimmunoresponsive cells comprising the above-described CARs. In certainembodiments, the isolated immunoresponsive cell is transduced with theCAR, for example, the CAR is constitutively expressed on the surface ofthe immunoresponsive cell. In certain embodiments, the isolatedimmunoresponsive cell is further transduced with at least oneco-stimulatory ligand such that the immunoresponsive cell expresses theat least one co-stimulatory ligand. In certain embodiments, the at leastone co-stimulatory ligand is selected from the group consisting of4-1BBL, CD80, CD86, CD70, OX40L, CD48, TNFRSF14, and combinationsthereof. In certain embodiments, the isolated immunoresponsive cell isfurther transduced with at least one cytokine such that theimmunoresponsive cell secrets the at least one cytokine. In certainembodiments, the at least cytokine is selected from the group consistingof IL-2, IL-3, IL-6, IL-7, IL-11, IL-12, IL-15, IL-17, IL-21, andcombinations thereof. In certain embodiments, the isolatedimmunoresponsive cell is selected from the group consisting of a T cell,a Natural Killer (NK) cell, a cytotoxic T lymphocyte (CTL), a regulatoryT cell, a human embryonic stem cell, a lymphoid progenitor cell, a Tcell-precursor cell, and a pluripotent stem cell from which lymphoidcells may be differentiated. In certain embodiments, theimmunoresponsive cell is a T cell.

The presently disclosed subject matter further provides nucleic acidmolecules encoding the presently disclosed CARs, vectors comprising thenucleic acid molecules, and host cells expressing such nucleic acidmolecules. In certain embodiments, the nucleic acid molecule comprisesnucleic acids having the sequence set forth in SEQ ID NO:207 In certainembodiments, the nucleic acid molecule comprises nucleic acids havingthe sequence set forth in SEQ ID NO:208. In certain embodiments, thenucleic acid molecule comprises nucleic acids having the sequence setforth in SEQ ID NO:209. In certain embodiments, the nucleic acidmolecule comprises nucleic acids having the sequence set forth in SEQ IDNO:229. In certain embodiments, the nucleic acid molecule comprisesnucleic acids having the sequence set forth in SEQ ID NO:230. In certainembodiments, the nucleic acid molecule comprises nucleic acids havingthe sequence set forth in SEQ ID NO:231. In certain embodiments, thenucleic acid molecule comprises nucleic acids having the sequence setforth in SEQ ID NO:232. In certain embodiments, the nucleic acidmolecule comprises nucleic acids having the sequence set forth in SEQ IDNO:233. In certain embodiments, the nucleic acid molecule comprisesnucleic acids having the sequence set forth in SEQ ID NO:234. In certainembodiments, the nucleic acid molecule comprises nucleic acids havingthe sequence set forth in SEQ ID NO:235. In certain embodiments, thevector is a γ-retroviral vector. In certain embodiments, the host cellis a T cell.

Furthermore, the presently disclosed subject matter provides methods ofusing the above-described immunoresponsive cell for reducing tumorburden in a subject. For example, the presently disclosed subject matterprovides methods of reducing tumor burden in a subject, where the methodcomprises administering an effective amount of the presently disclosedimmunoresponsive cell to the subject, thereby inducing tumor cell deathin the subject. In certain embodiments, the method reduces the number oftumor cells. In another embodiment, the method reduces the tumor size.In yet another embodiment, the method eradicates the tumor in thesubject. In certain embodiments, the tumor is selected from the groupconsisting of multiple myeloma, Non-Hodgkin Lymphoma, Hodgkin Lymphoma,Chronic Lymphocytic Leukemia (CLL), glioblastoma, and Waldenstrom'sMacroglobulinemia. In certain embodiments, the tumor is multiplemyeloma. In certain embodiments, the subject is a human. In certainembodiments, the immunoresponsive cell is a T cell.

Furthermore, the presently disclosed subject matter provides methods ofusing the above-described immunoresponsive cell for increasing orlengthening survival of a subject having neoplasia. For example, thepresently disclosed subject matter provides methods of increasing orlengthening survival of a subject having neoplasia, where the methodcomprises administering an effective amount of the presently disclosedimmunoresponsive cell to the subject, thereby increasing or lengtheningsurvival of the subject. In certain embodiments, the neoplasia isselected from the group consisting of multiple myeloma, Non-HodgkinLymphoma, Hodgkin Lymphoma, Chronic Lymphocytic Leukemia (CLL),glioblastoma, and Waldenstrom's Macroglobulinemia. In certainembodiments, the neoplasia is multiple myeloma. In certain embodiments,the method reduces or eradicates tumor burden in the subject.

The presently disclosed subject matter also provides methods forproducing an immunoresponsive cell that binds to BCMA. In onenon-limiting example, the method comprises introducing into theimmunoresponsive cell a nucleic acid sequence that encodes a chimericantigen receptor (CAR), which comprises an extracellular antigen-bindingdomain, a transmembrane domain and an intracellular domain, wherein theextracellular antigen-binding domain comprises a human scFv thatspecifically binds to BCMA.

The presently disclosed subject matter further provides pharmaceuticalcompositions comprising an effective amount of the presently disclosedimmunoresponsive cells and a pharmaceutically acceptable excipient. Incertain embodiments, the pharmaceutical compositions are for treating aneoplasia. In certain embodiments, the neoplasia is selected from thegroup consisting of multiple myeloma, Non-Hodgkin Lymphoma, HodgkinLymphoma, Chronic Lymphocytic Leukemia (CLL), glioblastoma, andWaldenstrom's Macroglobulinemia. In certain embodiments, the neoplasiais multiple myeloma.

The presently disclosed subject matter further provides kits fortreating a neoplasia, comprising the presently disclosedimmunoresponsive cells. In certain embodiments, the kit further includewritten instructions for using the immunoresponsive cell for treating aneoplasia. In certain embodiments, the neoplasia is selected from thegroup consisting of multiple myeloma, Non-Hodgkin Lymphoma, HodgkinLymphoma, Chronic Lymphocytic Leukemia (CLL), glioblastoma, andWaldenstrom's Macroglobulinemia. In certain embodiments, the neoplasiais multiple myeloma.

BRIEF DESCRIPTION OF THE FIGURES

The following Detailed Description, given by way of example, but notintended to limit the invention to specific embodiments described, maybe understood in conjunction with the accompanying drawings.

FIG. 1 shows a chimeric antigen receptor targeting BCMA in accordancewith one non-limiting embodiment of the presently disclosed subjectmatter.

FIGS. 2A-2D depict the human BCMA expression in normal tissues and humancancer cell lines.

FIG. 3 depicts expression of the presently disclosed BCMA CAR on human Tcells.

FIG. 4 depicts the cross-reacting activity of human scFv's targetinghuman BCMA with mouse BCMA.

FIG. 5 depicts the killing activity of the presently disclosed BCMA for3T3 cells overexpressing BCMA.

FIG. 6 depicts the killing activity of the presently disclosed BCMA fora human multiple myeloma cell line.

FIG. 7 shows a chimeric antigen receptor targeting BCMA in accordancewith one non-limiting embodiment of the presently disclosed subjectmatter.

FIG. 8 depicts a nucleic acid molecule that encodes a BCMA-targeted CARin accordance with one non-limiting embodiment of the presentlydisclosed subject matter.

FIG. 9 depicts a nucleic acid molecule that encodes a BCMA-targeted CARin accordance with one non-limiting embodiment of the presentlydisclosed subject matter.

FIG. 10 depicts a nucleic acid molecule that encodes a BCMA-targeted CARin accordance with one non-limiting embodiment of the presentlydisclosed subject matter.

FIG. 11 depicts a nucleic acid molecule that encodes a BCMA-targeted CARin accordance with one non-limiting embodiment of the presentlydisclosed subject matter.

FIG. 12 depicts a nucleic acid molecule that encodes a BCMA-targeted CARin accordance with one non-limiting embodiment of the presentlydisclosed subject matter.

FIG. 13 depicts the cytotoxicity of BCMA targeted CAR T cells for humanmultiple myeloma cell lines.

FIG. 14 depicts induction of cytokine secretion of BCMA targeted CAR Tcells.

FIG. 15 depicts anti-tumor activity of BCMA targeted CAR T cells.

FIGS. 16A and 16B depict the killing activity of BCMA targeted CAR Tcells. (A) Shows the percent of GFP⁺ tumor line at time 0. (B) Shows thekilling the percent of GFP⁺ tumor line at time 36 hours.

FIG. 17 depicts epitope mapping of ET140-3.

FIG. 18 depicts epitope mapping of ET140-24.

FIG. 19 depicts epitope mapping of ET140-54.

FIG. 20 depicts epitope mapping of ET140-3, ET140-24, and ET140-54.

FIG. 21 depicts ELISA screening data of ET140-3, ET140-24, ET140-37,ET140-40, and ET140-54.

FIGS. 22A-22D depict FCAS screening data of ET140-3 (FIG. 22B), ET140-24(FIG. 22C), ET140-37 (FIG. 22C), ET140-40 (FIG. 22D), and ET140-54 (FIG.22D).

DETAILED DESCRIPTION OF THE INVENTION

The presently disclosed subject matter generally provides chimericantigen receptors (CARs) targeting BCMA. In one non-limiting example,the CAR comprises an extracellular antigen-binding domain, atransmembrane domain and an intracellular domain, where theextracellular antigen-binding domain specifically binds to BCMA. Thepresently disclosed subject matter also provides immunoresponsive cells(e.g., T cell, a Natural Killer (NK) cell, a cytotoxic T lymphocyte(CTL), a regulatory T cell, a human embryonic stem cell, a lymphoidprogenitor cell, a T cell-precursor cell, and a pluripotent stem cellfrom which lymphoid cells may be differentiated) expressing theBCMA-targeted CARs, and methods of using such immunoresponsive cells fortreating a tumor, e.g., multiple myeloma.

I. Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the meaning commonly understood by a person skilled in the art towhich this invention belongs. The following references provide one ofskill with a general definition of many of the terms used in thisinvention: Singleton et al., Dictionary of Microbiology and MolecularBiology (2nd ed. 1994); The Cambridge Dictionary of Science andTechnology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R.Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, TheHarper Collins Dictionary of Biology (1991). As used herein, thefollowing terms have the meanings ascribed to them below, unlessspecified otherwise.

As used herein, the term “about” or “approximately” means within anacceptable error range for the particular value as determined by one ofordinary skill in the art, which will depend in part on how the value ismeasured or determined, i.e., the limitations of the measurement system.For example, “about” can mean within 3 or more than 3 standarddeviations, per the practice in the art. Alternatively, “about” can meana range of up to 20%, preferably up to 10%, more preferably up to 5%,and more preferably still up to 1% of a given value. Alternatively,particularly with respect to biological systems or processes, the termcan mean within an order of magnitude, preferably within 5-fold, andmore preferably within 2-fold, of a value.

As used herein, the term “cell population” refers to a group of at leasttwo cells expressing similar or different phenotypes. In non-limitingexamples, a cell population can include at least about 10, at leastabout 100, at least about 200, at least about 300, at least about 400,at least about 500, at least about 600, at least about 700, at leastabout 800, at least about 900, at least about 1000 cells expressingsimilar or different phenotypes.

As used herein, the term “antibody” means not only intact antibodymolecules, but also fragments of antibody molecules that retainimmunogen-binding ability. Such fragments are also well known in the artand are regularly employed both in vitro and in vivo. Accordingly, asused herein, the term “antibody” means not only intact immunoglobulinmolecules but also the well-known active fragments F(ab′)2, and Fab.F(ab′)2, and Fab fragments that lack the Fe fragment of intact antibody,clear more rapidly from the circulation, and may have less non-specifictissue binding of an intact antibody (Wahl et al., J. Nucl. Med.24:316-325 (1983). The antibodies of the invention comprise whole nativeantibodies, bispecific antibodies; chimeric antibodies; Fab, Fab′,single chain V region fragments (scFv), fusion polypeptides, andunconventional antibodies.

As used herein, the term “single-chain variable fragment” or “scFv” is afusion protein of the variable regions of the heavy (V_(H)) and lightchains (V_(L)) of an immunoglobulin (e.g., mouse or human) covalentlylinked to form a V_(H)::VL heterodimer. The heavy (V_(H)) and lightchains (V_(L)) are either joined directly or joined by apeptide-encoding linker (e.g., 10, 15, 20, 25 amino acids), whichconnects the N-terminus of the V_(H) with the C-terminus of the V_(L),or the C-terminus of the V_(H) with the N-terminus of the V_(L). Thelinker is usually rich in glycine for flexibility, as well as serine orthreonine for solubility. The linker can link the heavy chain variableregion and the light chain variable region of the extracellularantigen-binding domain. Non-limiting examples of linkers are disclosedin Shen et al., Anal. Chem. 80(6):1910-1917 (2008) and WO 2014/087010,the contents of which are hereby incorporated by reference in theirentireties. In certain embodiments, the linker is a G45 linker.

In a non-limiting example, the linker comprises amino acids having thesequence set forth in SEQ ID NO:210 as provided below.

[SEQ ID NO: 210] GGGGSGGGGSGGGGS.

In certain embodiments, the nucleic acid sequence encoding the aminoacid sequence of SEQ ID NO:210 is set forth in SEQ ID NO:211, which isprovided below:

[SEQ ID NO: 211] GGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCT.

In one non-limiting example, the linker comprises amino acids having thesequence set forth in SEQ ID NO:69 as provided below.

[SEQ ID NO: 69] SRGGGGSGGGGSGGGGSLEMA

In certain embodiments, the nucleic acid sequence encoding the aminoacid sequence of SEQ ID NO:69 is set forth in SEQ ID NO:70, which isprovided below:

[SEQ ID NO: 70] tctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggcc

Despite removal of the constant regions and the introduction of alinker, scFv proteins retain the specificity of the originalimmunoglobulin. Single chain Fv polypeptide antibodies can be expressedfrom a nucleic acid comprising V_(H)- and V_(L)-encoding sequences asdescribed by Huston, et al. (Proc. Nat. Acad. Sci. USA, 85:5879-5883,1988). See, also, U.S. Pat. Nos. 5,091,513, 5,132,405 and 4,956,778; andU.S. Patent Publication Nos. 20050196754 and 20050196754. AntagonisticscFvs having inhibitory activity have been described (see, e.g., Zhao etal., Hyrbidoma (Larchmt) 2008 27(6):455-51; Peter et al., J CachexiaSarcopenia Muscle 2012 Aug. 12; Shieh et al., J Imunol 2009183(4):2277-85; Giomarelli et al., Thromb Haemost 2007 97(6):955-63;Fife eta., J Clin Inst 2006 116(8):2252-61; Brocks et al.,Immunotechnology 1997 3(3):173-84; Moosmayer et al., Ther Immunol 19952(10:31-40). Agonistic scFvs having stimulatory activity have beendescribed (see, e.g., Peter et al., J Bioi Chern 2003 25278(38):36740-7;Xie et al., Nat Biotech 1997 15(8):768-71; Ledbetter et al., Crit RevImmunol 1997 17(5-6):427-55; Ho et al., BioChim Biophys Acta 20031638(3):257-66).

As used herein, “F(ab)” refers to a fragment of an antibody structurethat binds to an antigen but is monovalent and does not have a Fcportion, for example, an antibody digested by the enzyme papain yieldstwo F(ab) fragments and an Fc fragment (e.g., a heavy (H) chain constantregion; Fc region that does not bind to an antigen).

As used herein, “F(ab′)2” refers to an antibody fragment generated bypepsin digestion of whole IgG antibodies, wherein this fragment has twoantigen binding (ab′) (bivalent) regions, wherein each (ab′) regioncomprises two separate amino acid chains, a part of a H chain and alight (L) chain linked by an S—S bond for binding an antigen and wherethe remaining H chain portions are linked together. A “F(ab′)2” fragmentcan be split into two individual Fab′ fragments.

As used herein, the term “vector” refers to any genetic element, such asa plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc.,which is capable of replication when associated with the proper controlelements and which can transfer gene sequences into cells. Thus, theterm includes cloning and expression vehicles, as well as viral vectorsand plasmid vectors.

As used herein, the term “expression vector” refers to a recombinantnucleic acid sequence, i.e. recombinant DNA molecule, containing adesired coding sequence and appropriate nucleic acid sequences necessaryfor the expression of the operably linked coding sequence in aparticular host organism. Nucleic acid sequences necessary forexpression in prokaryotes usually include a promoter, an operator(optional), and a ribosome binding site, often along with othersequences. Eukaryotic cells are known to utilize promoters, enhancers,and termination and polyadenylation signals.

As used herein, “CDRs” are defined as the complementarity determiningregion amino acid sequences of an antibody which are the hypervariableregions of immunoglobulin heavy and light chains. See, e.g., Kabat etal., Sequences of Proteins of Immunological Interest, 4th U. S.Department of Health and Human Services, National Institutes of Health(1987). Generally, antibodies comprise three heavy chain and three lightchain CDRs or CDR regions in the variable region. CDRs provide themajority of contact residues for the binding of the antibody to theantigen or epitope. In certain embodiments, the CDRs regions aredelineated using the Kabat system (Kabat, E. A., et al. (1991) Sequencesof Proteins of Immunological Interest, Fifth Edition, U.S. Department ofHealth and Human Services, NIH Publication No. 91-3242).

As used herein, the term “affinity” is meant a measure of bindingstrength. Without being bound to theory, affinity depends on thecloseness of stereochemical fit between antibody combining sites andantigen determinants, on the size of the area of contact between them,and on the distribution of charged and hydrophobic groups. Affinity alsoincludes the term “avidity,” which refers to the strength of theantigen-antibody bond after formation of reversible complexes. Methodsfor calculating the affinity of an antibody for an antigen are known inthe art, comprising use of binding experiments to calculate affinity.Antibody activity in functional assays (e.g., flow cytometry assay) isalso reflective of antibody affinity. Antibodies and affinities can bephenotypically characterized and compared using functional assays (e.g.,flow cytometry assay).

Nucleic acid molecules useful in the methods of the invention includeany nucleic acid molecule that encodes a polypeptide of the invention ora fragment thereof. Such nucleic acid molecules need not be 100%identical with an endogenous nucleic acid sequence, but will typicallyexhibit substantial identity. Polynucleotides having “substantialidentity” to an endogenous sequence are typically capable of hybridizingwith at least one strand of a double-stranded nucleic acid molecule. By“hybridize” is meant pair to form a double-stranded molecule betweencomplementary polynucleotide sequences (e.g., a gene described herein),or portions thereof, under various conditions of stringency. (See, e.g.,Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399; Kimmel, A.R. (1987) Methods Enzymol. 152:507).

For example, stringent salt concentration will ordinarily be less thanabout 750 mM NaCl and 75 mM trisodium citrate, preferably less thanabout 500 mM NaCl and 50 mM trisodium citrate, and more preferably lessthan about 250 mM NaCl and 25 mM trisodium citrate. Low stringencyhybridization can be obtained in the absence of organic solvent, e.g.,formamide, while high stringency hybridization can be obtained in thepresence of at least about 35% formamide, and more preferably at leastabout 50% formamide. Stringent temperature conditions will ordinarilyinclude temperatures of at least about 30° C., more preferably of atleast about 37° C., and most preferably of at least about 42° C. Varyingadditional parameters, such as hybridization time, the concentration ofdetergent, e.g., sodium dodecyl sulfate (SDS), and the inclusion orexclusion of carrier DNA, are well known to those skilled in the art.Various levels of stringency are accomplished by combining these variousconditions as needed. In a preferred: embodiment, hybridization willoccur at 30° C. in 750 mM NaCl, 75 mM trisodium citrate, and 1% SDS. Ina more preferred embodiment, hybridization will occur at 37° C. in 500mM NaCl, 50 mM trisodium citrate, 1% SDS, 35% formamide, and 100 μg/mldenatured salmon sperm DNA (ssDNA). In a most preferred embodiment,hybridization will occur at 42° C. in 250 mM NaCl, 25 mM trisodiumcitrate, 1% SDS, 50% formamide, and 200 μg/ml ssDNA. Useful variationson these conditions will be readily apparent to those skilled in theart.

For most applications, washing steps that follow hybridization will alsovary in stringency. Wash stringency conditions can be defined by saltconcentration and by temperature. As above, wash stringency can beincreased by decreasing salt concentration or by increasing temperature.For example, stringent salt concentration for the wash steps willpreferably be less than about 30 mM NaCl and 3 mM trisodium citrate, andmost preferably less than about 15 mM NaCl and 1.5 mM trisodium citrate.Stringent temperature conditions for the wash steps will ordinarilyinclude a temperature of at least about 25° C., more preferably of atleast about 42° C., and even more preferably of at least about 68° C. Ina preferred embodiment, wash steps will occur at 25° C. in 30 mM NaCl, 3mM trisodium citrate, and 0.1% SDS. In a more preferred embodiment, washsteps will occur at 42° C. in 15 mM NaCl, 1.5 mM trisodium citrate, and0.1% SDS. In a more preferred embodiment, wash steps will occur at 68°C. in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. Additionalvariations on these conditions will be readily apparent to those skilledin the art. Hybridization techniques are well known to those skilled inthe art and are described, for example, in Benton and Davis (Science196:180, 1977); Grunstein and Rogness (Proc. Natl. Acad. Sci., USA72:3961, 1975); Ausubel et al. (Current Protocols in Molecular Biology,Wiley Interscience, New York, 2001); Berger and Kimmel (Guide toMolecular Cloning Techniques, 1987, Academic Press, New York); andSambrook et al., Molecular Cloning: A Laboratory Manual, Cold SpringHarbor Laboratory Press, New York.

By “substantially identical” is meant a polypeptide or nucleic acidmolecule exhibiting at least 50% identity to a reference amino acidsequence (for example, any one of the amino acid sequences describedherein) or nucleic acid sequence (for example, any one of the nucleicacid sequences described herein). Preferably, such a sequence is atleast 60%, more preferably 80% or 85%, and more preferably 90%, 95% oreven 99% identical at the amino acid level or nucleic acid to thesequence used for comparison.

Sequence identity is typically measured using sequence analysis software(for example, Sequence Analysis Software Package of the GeneticsComputer Group, University of Wisconsin Biotechnology Center, 1710University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, orPILEUP/PRETTYBOX programs). Such software matches identical or similarsequences by assigning degrees of homology to various substitutions,deletions, and/or other modifications. In an exemplary approach todetermining the degree of identity, a BLAST program may be used, with aprobability score between e-3 and e-100 indicating a closely relatedsequence.

As used herein, the term “analog” refers to a structurally relatedpolypeptide or nucleic acid molecule having the function of a referencepolypeptide or nucleic acid molecule.

As used herein, the term “ligand” refers to a molecule that binds to areceptor. In particular, the ligand binds a receptor on another cell,allowing for cell-to-cell recognition and/or interaction.

As used herein, the term “disease” refers to any condition or disorderthat damages or interferes with the normal function of a cell, tissue,or organ. Examples of diseases include neoplasia or pathogen infectionof cell.

As used herein, the term “effective amount” refers to an amountsufficient to have a therapeutic effect. In certain embodiments, an“effective amount” is an amount sufficient to arrest, ameliorate, orinhibit the continued proliferation, growth, or metastasis (e.g.,invasion, or migration) of a neoplasia.

As used herein, the term “heterologous nucleic acid molecule orpolypeptide” refers to a nucleic acid molecule (e.g., a cDNA, DNA or RNAmolecule) or polypeptide that is not normally present in a cell orsample obtained from a cell. This nucleic acid may be from anotherorganism, or it may be, for example, an mRNA molecule that is notnormally expressed in a cell or sample.

As used herein, the term “immunoresponsive cell” refers to a cell thatfunctions in an immune response or a progenitor, or progeny thereof.

As used herein, the term “modulate” refers positively or negativelyalter. Exemplary modulations include an about 1%, about 2%, about 5%,about 10%, about 25%, about 50%, about 75%, or about 100% change.

As used herein, the term “increase” refers to alter positively by atleast about 5%, including, but not limited to, alter positively by about5%, by about 10%, by about 25%, by about 30%, by about 50%, by about75%, or by about 100%.

As used herein, the term “reduce” refers to alter negatively by at leastabout 5% including, but not limited to, alter negatively by about 5%, byabout 10%, by about 25%, by about 30%, by about 50%, by about 75%, or byabout 100%.

As used herein, the term “isolated cell” refers to a cell that isseparated from the molecular and/or cellular components that naturallyaccompany the cell.

As used herein, the term “isolated,” “purified,” or “biologically pure”refers to material that is free to varying degrees from components whichnormally accompany it as found in its native state. “Isolate” denotes adegree of separation from original source or surroundings. “Purify”denotes a degree of separation that is higher than isolation. A“purified” or “biologically pure” protein is sufficiently free of othermaterials such that any impurities do not materially affect thebiological properties of the protein or cause other adverseconsequences. That is, a nucleic acid or peptide of this invention ispurified if it is substantially free of cellular material, viralmaterial, or culture medium when produced by recombinant DNA techniques,or chemical precursors or other chemicals when chemically synthesized.Purity and homogeneity are typically determined using analyticalchemistry techniques, for example, polyacrylamide gel electrophoresis orhigh performance liquid chromatography. The term “purified” can denotethat a nucleic acid or protein gives rise to essentially one band in anelectrophoretic gel. For a protein that can be subjected tomodifications, for example, phosphorylation or glycosylation, differentmodifications may give rise to different isolated proteins, which can beseparately purified.

As used herein, the term “secreted” is meant a polypeptide that isreleased from a cell via the secretory pathway through the endoplasmicreticulum, Golgi apparatus, and as a vesicle that transiently fuses atthe cell plasma membrane, releasing the proteins outside of the cell.

As used herein, the term “specifically binds” or “specifically binds to”or “specifically target” is meant a polypeptide or fragment thereof thatrecognizes and binds a biological molecule of interest (e.g., apolypeptide), but which does not substantially recognize and bind othermolecules in a sample, for example, a biological sample, which naturallyincludes a polypeptide of the invention.

As used herein, the term “treating” or “treatment” refers to clinicalintervention in an attempt to alter the disease course of the individualor cell being treated, and can be performed either for prophylaxis orduring the course of clinical pathology. Therapeutic effects oftreatment include, without limitation, preventing occurrence orrecurrence of disease, alleviation of symptoms, diminishment of anydirect or indirect pathological consequences of the disease, preventingmetastases, decreasing the rate of disease progression, amelioration orpalliation of the disease state, and remission or improved prognosis. Bypreventing progression of a disease or disorder, a treatment can preventdeterioration due to a disorder in an affected or diagnosed subject or asubject suspected of having the disorder, but also a treatment mayprevent the onset of the disorder or a symptom of the disorder in asubject at risk for the disorder or suspected of having the disorder.

As used herein, the term “subject” refers to any animal (e.g., amammal), including, but not limited to, humans, non-human primates,rodents, and the like (e.g., which is to be the recipient of aparticular treatment, or from whom cells are harvested).

II. B Cell Maturation Antigen (BCMA)

BCMA is an ideal target for Adoptive T cell therapy (e.g., CAR therapy)as BCMA is involved in B cell differentiation and signaling and is knownto be expressed on non-malignant differentiated B cells and plasmacells. While there might be risk of inducing a B cell aplasia, B cellaplasias induced by the CD19 CAR have been remarkably well tolerated.Several groups have confirmed BCMA multiple myeloma (MM) surfaceexpression, with one group finding it as an alternative to CD138 as aFACS marker for malignant plasma cells from fresh or frozen patient bonemarrow samples with mean relative mean fluorescence intensity (MFI)between 9-16 (n=35).^(1,2)

In certain non-limiting embodiments, BCMA is human BCMA having the aminoacid sequence set forth in SEQ ID NO:71, or fragments thereof.

SEQ ID NO:71 is provided below:

[SEQ ID NO: 71] MLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNAILWTCLGLSLIISLAVFVLMFLLRKINSEPLKDEFKNTGSGLLGMANIDLEKSRTGDEIILPRGLEYTVEECTCEDCIKSKPKVDSDHCFPLPAMEEGATILVTTKTNDYCKSLPAALSATEIEKSISAR

III. Chimeric Antigen Receptor (CAR)

Chimeric antigen receptors (CARs) are engineered receptors, which graftor confer a specificity of interest onto an immune effector cell. CARscan be used to graft the specificity of a monoclonal antibody onto a Tcell; with transfer of their coding sequence facilitated by retroviralvectors.

There are three generations of CARs. “First generation” CARs aretypically composed of an extracellular antigen binding domain (e.g., asingle-chain variable fragments (scFv)) fused to a transmembrane domain,fused to cytoplasmic/intracellular domain of the T cell receptor chain.“First generation” CARs typically have the intracellular domain from theCD3ξ− chain, which is the primary transmitter of signals from endogenousTCRs. “First generation” CARs can provide de novo antigen recognitionand cause activation of both CD4⁺ and CD8⁺ T cells through their CD3ζchain signaling domain in a single fusion molecule, independent ofHLA-mediated antigen presentation. “Second generation” CARs addintracellular domains from various co-stimulatory molecules (e.g., CD28,4-1BB, ICOS, OX40) to the cytoplasmic tail of the CAR to provideadditional signals to the T cell. “Second generation” CARs comprisethose that provide both co-stimulation (e.g., CD28 or 4-1BB) andactivation (CD3ζ). Preclinical studies have indicated that “SecondGeneration” CARs can improve the anti-tumor activity of T cells. Forexample, robust efficacy of “Second Generation” CAR modified T cells wasdemonstrated in clinical trials targeting the CD19 molecule in patientswith chronic lymphoblastic leukemia (CLL) and acute lymphoblasticleukemia (ALL). “Third generation” CARs comprise those that providemultiple co-stimulation (e.g., CD28 and 4-1BB) and activation (CD3ζ).

In accordance with the presently disclosed subject matter, the CARscomprise an extracellular antigen-binding domain, a transmembrane domainand an intracellular domain, where the extracellular antigen-bindingdomain binds to BCMA. In certain embodiments, the extracellularantigen-binding domain is a scFv. In certain embodiments, theextracellular antigen-binding domain is a Fab, which is optionallycrosslinked. In a certain embodiments, the extracellular binding domainis a F(ab)₂. In certain embodiments, any of the foregoing molecules maybe comprised in a fusion protein with a heterologous sequence to formthe extracellular antigen-binding domain. In one specific non-limitingembodiment, the extracellular antigen-binding domain comprises a humanscFv that binds specifically to human BCMA.

In certain non-limiting embodiments, the extracellular antigen-bindingdomain of a CAR has a high binding specificity as well as high bindingaffinity to the BCMA. For example, in such embodiments, theextracellular antigen-binding domain of the CAR (embodied, for example,in a human scFv or an analog thereof) binds to BCMA with a dissociationconstant (K_(D)) of about 3×10⁻⁶ M or less. In certain embodiments, theK_(D) is about 1×10⁻⁶ M or less, about 1×10⁻⁷ M or less, about 1×10⁻⁸Mor less, or about 1×10⁻⁹M or less, about 1×10⁻¹⁰ M or less, or about1×10⁻¹¹M or less.

In certain embodiments, the K_(D) is about 1×10⁻⁸ M or less. In certainembodiments, the K_(D) is from about 1×10⁻¹¹M to about 3×10⁻⁶ M, such asfrom about 1×10⁻¹¹M to about 1×10⁻¹⁰ M, from about 1×10⁻¹° M to about1×10⁻⁹M, from about 1×10⁻⁹M to about 1×10⁻⁸M, from about 1×10⁻⁸ M toabout 1×10⁻⁷ M, or from about 1×10⁻⁷ M to about 1×10⁻⁶ M, or from about1×10⁻⁶ M to about 3×10⁻⁶ M. In certain embodiments, the K_(D) is fromabout 1×10⁻⁹M to about 1×10⁻⁸ M. In certain embodiments, the K_(D) isfrom about 1×10⁻⁹ M to about 1.5×10⁻⁹ M. In certain embodiments, theK_(D) is about 1.2×10⁻⁹M. In certain embodiments, the K_(D) is fromabout 4×10⁻⁹ M to about 5×10⁻⁹ M. In certain embodiments, the K_(D) isabout 5×10⁻⁹ M. In certain embodiments, the K_(D) is about 4.8×10⁻⁹ M.In certain embodiments, the K_(D) is from about 8×10⁻⁹M to about 9×10⁻⁹M. In certain embodiments, the K_(D) is about 8×10⁻⁹M. In certainembodiments, the K_(D) is about 8.1×10⁻⁹M.

Binding of the extracellular antigen-binding domain (embodiment, forexample, in a human scFv or an analog thereof) of a presently disclosedCAR to BCMA can be confirmed by, for example, enzyme-linkedimmunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis,bioassay (e.g., growth inhibition), or Western Blot assay. Each of theseassays generally detect the presence of protein-antibody complexes ofparticular interest by employing a labeled reagent (e.g., an antibody,or a scFv) specific for the complex of interest. For example, the scFvcan be radioactively labeled and used in a radioimmunoassay (RIA) (see,for example, Weintraub, B., Principles of Radioimmunoassays, SeventhTraining Course on Radioligand Assay Techniques, The Endocrine Society,March, 1986, which is incorporated by reference herein). The radioactiveisotope can be detected by such means as the use of a γ counter or ascintillation counter or by autoradiography. In certain embodiments, theBCMA-targeted extracellular antigen-binding domain is labeled with afluorescent marker. Non-limiting examples of fluorescent markers includegreen fluorescent protein (GFP), blue fluorescent protein (e.g., EBFP,EBFP2, Azurite, and mKalamal), cyan fluorescent protein (e.g., ECFP,Cerulean, and CyPet), and yellow fluorescent protein (e.g., YFP,Citrine, Venus, and YPet). In certain embodiments, the BCMA-targetedhuman scFv is labeled with GFP.

In certain embodiments, the extracellular antigen-binding domain of apresently disclosed CAR comprises a single-chain variable fragment(scFv). In one specific non-limiting embodiment, the extracellularantigen-binding domain of a presently disclosed CAR comprises a humanscFv that specifically binds to human BCMA. In certain embodiments, thescFv are identified by screening scFv phage library with BCMA-Fc fusionprotein.

Extracellular Antigen Binding Domain of a CAR

In certain embodiments, the extracellular antigen-binding domain (e.g.,human scFv) comprises a heavy chain variable region comprising aminoacids having a sequence selected from the group consisting of: SEQ IDNOS:1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, and 65.The nucleic acid sequences encoding the amino acid sequence of SEQ IDNOS:1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, and 65are 3, 7, 11, 15, 19, 23, 27, 31, 35, 39, 43, 47, 51, 55, 59, 63, and67, respectively. In certain embodiments, the extracellularantigen-binding domain (e.g., human scFv) comprises a light chainvariable region comprising amino acids having a sequence selected fromthe group consisting of SEQ ID NOS: 2, 6, 10, 14, 18, 22, 26, 30, 34,38, 42, 46, 50, 54, 58, 62, and 66. The nucleic acid sequences encodingthe amino acid sequence of SEQ ID NOS: 2, 6, 10, 14, 18, 22, 26, 30, 34,38, 42, 46, 50, 54, 58, 62, and 66 are 4, 8, 12, 16, 20, 24, 28, 32, 36,40, 44, 48, 52, 56, 60, 64, and 68, respectively. The sequences of SEQID NOS:1-68 are described in the following Tables 1-17.

In certain embodiments, the extracellular antigen-binding domain (e.g.,scFv) comprises heavy and light chain variable regions comprising aminoacid sequences that are homologous to the amino acid sequences describedherein and as disclosed in Tables 1-17. For example, and not by way oflimitation, the extracellular antigen-binding domain (e.g., scFv)comprises a heavy chain variable region comprising an amino acidsequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homologous to anamino acid sequence selected from the group consisting of SEQ ID NOS: 1,5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, and 65.

In certain embodiments, the extracellular antigen-binding domain (e.g.,scFv) comprises a light chain variable region comprising an amino acidsequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homologous to anamino acid sequence selected from the group consisting of SEQ ID NOS: 2,6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, and 66 are 4,8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, and 68.

In certain embodiments, the extracellular antigen-binding domain (e.g.,scFv) comprises (a) a heavy chain variable region comprising an aminoacid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homologousto an amino acid sequence selected from the group consisting of SEQ IDNOS: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, and65; and (b) a light chain variable region comprising an amino acidsequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homologous to anamino acid sequence selected from the group consisting of SEQ ID NOS: 2,6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, and 66 are 4,8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, and 68.

The presently disclosed subject matter further provides extracellularantigen-binding domains (e.g., scFv) that comprise heavy chain variableregion and light chain variable region CDRs, e.g., CDR1s, CDR2s andCDR3s, as disclosed herein in Tables 1-17. The CDR regions aredelineated using the Kabat system (Kabat, E. A., et al. (1991) Sequencesof Proteins of Immunological Interest, Fifth Edition, U.S. Department ofHealth and Human Services, NIH Publication No. 91-3242). The presentlydisclosed subject matter further provides extracellular antigen-bindingdomains (e.g., scFv) that comprise conservative modifications of theantibody sequences disclosed herein. For example, and not by way oflimitation, an extracellular antigen-binding domains (e.g., scFv) of thepresently disclosed subject matter comprises a heavy chain variableregion comprising CDR1, CDR2 and CDR3 sequences and a light chainvariable region comprising CDR1, CDR2 and CDR3 sequences, wherein one ormore of these CDR sequences comprise specified amino acid sequencesdisclosed herein, or conservative modifications thereof, and wherein theextracellular antigen-binding domains retain the desired functionalproperties.

In certain embodiments, the presently disclosed subject matter providesan extracellular antigen-binding domain (e.g., scFv) comprising a heavychain variable region, wherein the heavy chain variable regioncomprises: (a) a CDR1 comprising an amino acid sequence selected fromthe group consisting of SEQ ID NOS: 89, 95, 101, 107, 113, 119, 125,131, 137, 143, 149, 155, 161, 167, 173, 179, and 185, and conservativemodifications thereof; (b) a CDR2 comprising an amino acid sequenceselected from the group consisting of SEQ ID NOS: 90, 96, 102, 108, 114,120, 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, and 186, andconservative modifications thereof; and (c) a CDR3 comprising an aminoacid sequence selected from the group consisting of SEQ ID NOS: 91, 97,103, 109, 115, 121, 127, 133, 139, 145, 151, 157, 163, 169, 175, 181,and 187, and conservative modifications thereof.

In certain embodiments, the extracellular antigen-binding domain (e.g.,scFv) comprises a light chain variable region, wherein the light chainvariable region comprises: (a) a CDR1 comprising an amino acid sequenceselected from the group consisting of SEQ ID NOS: 92, 98, 104, 110, 116,122, 128, 134, 140, 146, 152, 158, 164, 170, 176, 182, and 188, andconservative modifications thereof; (b) a CDR2 comprising an amino acidsequence selected from the group consisting of SEQ ID NOS: 93, 99, 105,111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183, and189, and conservative modifications thereof; and (c) a CDR3 comprisingan amino acid sequence selected from the group consisting of SEQ ID NOS:94, 100, 106, 112, 118, 124, 130, 136, 142, 148, 154, 160, 166, 172,178, 184, and 190, and conservative modifications thereof.

The presently disclosed subject matter provides an extracellularantigen-binding domain (e.g., scFv) comprising a heavy chain variableregion comprising CDR1, CDR2, and CDR3 sequences and a light chainvariable region comprising CDR1, CDR2, and CDR3 sequences, wherein: (a)the heavy chain variable region CDR3 comprises an amino acid sequenceselected from the group consisting of SEQ ID NOs: 91, 97, 103, 109, 115,121, 127, 133, 139, 145, 151, 157, 163, 169, 175, 181, and 187, andconservative modifications thereof; and (b) the light chain variableregion CDR3 comprises an amino acid sequence selected from the groupconsisting of SEQ ID NOS: 94, 100, 106, 112, 118, 124, 130, 136, 142,148, 154, 160, 166, 172, 178, 184, and 190, and conservativemodifications thereof; wherein the extracellular antigen-binding domainspecifically binds to a BCMA polypeptide (e.g., a human BCMApolypeptide). In certain embodiments, the heavy chain variable regionCDR2 comprises an amino acid sequence selected from the group consistingof SEQ ID NOs: 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185,191, 197, 203, 209, 215, 221, 227, 233, 239, 245, 251, 257, 263, 305,317, 329, 341, 353, 365, 377, and 389, and conservative modificationsthereof; and (b) the light chain variable region CDR2 comprises an aminoacid sequence selected from the group consisting of SEQ ID NOS: 93, 99,105, 111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183,and 189, and conservative modifications thereof; wherein theextracellular antigen-binding domain specifically binds to a BCMApolypeptide (e.g., a human BCMA polypeptide). In certain embodiments,the heavy chain variable region CDR1 comprises an amino acid sequenceselected from the group consisting of SEQ ID NOs: 89, 95, 101, 107, 113,119, 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, and 185, andconservative modifications thereof; and (b) the light chain variableregion CDR1 comprises an amino acid sequence selected from the groupconsisting of SEQ ID NOS: 92, 98, 104, 110, 116, 122, 128, 134, 140,146, 152, 158, 164, 170, 176, 182, and 188, and conservativemodifications thereof; wherein the extracellular antigen-binding domainspecifically binds to a BCMA polypeptide (e.g., a human BCMApolypeptide).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO: 72 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-192 scFv (also referred to as “ET140-42scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:1 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:2, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 1. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:1, as shownin Table 1. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(H) comprising amino acids having the sequence setforth in SEQ ID NO:1, as shown in Table 1. In certain embodiments, theextracellular antigen-binding domain comprises a V_(L) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:2, as shownin Table 1. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) comprising amino acids having the sequence setforth in SEQ ID NO:2, as shown in Table 1. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising aminoacids having the sequence set forth in SEQ ID NO:1 and a V_(L)comprising amino acids having the sequence set forth in SEQ ID NO:2, asshown in Table 1. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:89 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:90 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:91 or conservative modifications thereof, as shown inTable 1. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:92 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:93 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO:94 orconservative modifications thereof, as shown in Table 1. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:89 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:90 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:91 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:92 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:93 or conservative modifications thereof, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:94 or conservative modifications thereof, as shown in Table 1. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:89, a V_(H) CDR2 mprising amino acids having the sequence setforth in SEQ ID NO:90, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:91, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:92, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:93, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:94.

TABLE 1 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) VSSNSAAWN YRSKWYNARQGYSYYGYSDV [SEQ ID [SEQ ID [SEQ ID NO: 89] NO: 90] NO: 91] V_(L)SSNIGHND FDD AAWDGSLNAFV [SEQ ID [SEQ ID [SEQ ID NO: 92] NO: 93 NO: 94]Full QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSP V_(H)SRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARQGYSYYGYSDVWGQGTLVTVSS [SEQ ID NO: 1] DNACaggtacagctgcagcagtcaggtccaggactggtgaagccctcgcagaccctctcactcacctgtgccatctccggggacagtgtctctagcaacagtgctgcttggaactggatcaggcagtccccatcgagaggccttgagtggctgggaaggacatactacaggtccaagtggtataatgattatgcagtatctgtgaaaagtcgaataaccatcaacccagacacatccaagaaccagttctccctgcagctgaactctgtgactcccgaggacacggctgtgtattactgtgcgcgccagggttactcttactacggttactctgatgtttggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 3] FullQSVLTQPPSVSVAPRQRVTISCSGSSSNIGHNDVSWYQHLPGK V_(L)APRLLIYFDDLLPSGVSDRFSASKSGTSASLAISGLQSEDEADYYCAAWDGSLNAFVFGTGTKVTVLG [SEQ ID NO: 2] DNACagtctgtgctgactcagccaccctcggtgtctgtagcccccaggcagagggtcaccatctcgtgttctggaagcagctccaacatcggacataatgatgtaagctggtaccagcatctcccagggaaggctcccagactcctcatctattttgatgacctgctgccgtcaggggtctctgaccgattctctgcctccaagtctggcacctcagcctccctggccatcagtgggctccagtctgaggatgaggctgattattactgtgcagcatgggatggcagcctgaatgcctttgtcttcggaactgggaccaaggtcaccgtcctaggt [SEQ ID NO: 4] scFvQSVLTQPPSVSVAPRQRVTISCSGSSSNIGHNDVSWYQHLPGKAPRLLIYFDDLLPSGVSDRFSASKSGTSASLAISGLQSEDEADYYCAAWDGSLNAFVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARQGYSYYGYSDVWGQGTLVTVSS [SEQ ID NO: 72]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:73 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-197 scFv (also referred to as “ET140-47scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:5 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:6, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 2. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:5, as shownin Table 2. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(H) comprising amino acids having the sequence setforth in SEQ ID NO:5, as shown in Table 2. In certain embodiments, theextracellular antigen-binding domain comprises a V_(L) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:6, as shownin Table 2. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) comprising amino acids having the sequence setforth in SEQ ID NO:6, as shown in Table 2. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising aminoacids having the sequence set forth in SEQ ID NO:5 and a V_(L)comprising amino acids having the sequence set forth in SEQ ID NO:6, asshown in Table 2. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:95 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:96 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:97 or conservative modifications thereof, as shown inTable 2. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:98 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:99 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO:100 orconservative modifications thereof, as shown in Table 2. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:95 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:96 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:97 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:98 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:99 or conservative modifications thereof, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:100 or conservative modifications thereof, as shown in Table 2. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:95, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:96, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:97, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:98, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:99, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:100.

TABLE 2 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) VSSNSAAWN YRSKWYNARYGFSGSRFYDT [SEQ ID [SEQ ID [SEQ ID NO: 95] NO: 96] NO: 97] V_(L)SSNIGNNA FDD AAWDDSLNGYV [SEQ ID [SEQ ID [SEQ ID NO: 98] NO: 99]NO: 100] Full QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSP V_(H)SRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARYGFSGSRFYDTWGQGTLVTVSS [SEQ ID NO: 5] DNACaggtacagctgcagcagtcaggtccaggactggtgaagccctcgcagaccctctcactcacctgtgccatctccggggacagtgtctctagcaacagtgctgcttggaactggatcaggcagtccccatcgagaggccttgagtggctgggaaggacatactacaggtccaagtggtataatgattatgcagtatctgtgaaaagtcgaataaccatcaacccagacacatccaagaaccagttctccctgcagctgaactctgtgactcccgaggacacggctgtgtattactgtgcgcgctacggtttctctggttctcgtttctacgatacttggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 7] FullQPVLTQPPSVSEAPRQRVTISCSGSSSNIGNNAVNWYQQLPGK V_(L)APKLLIYFDDLLSSGVSDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGYVFGTGTKVTVLG [SEQ ID NO: 6] DNACagcctgtgctgactcagccaccctcggtgtctgaagcccccaggcagagggtcaccatctcctgttctggaagcagctccaacatcggaaataatgctgtaaactggtaccagcagctcccaggaaaggctcccaaactcctcatctattttgatgatctgctgtcctcaggggtctctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtgggctccagtctgaagatgaggctgattattactgtgcagcatgggatgacagcctgaatggttatgtcttcggaactgggaccaaggtcaccgtcctaggt [SEQ ID NO: 8] scFvQPVLTQPPSVSEAPRQRVTISCSGSSSNIGNNAVNWYQQLPGKAPKLLIYFDDLLSSGVSDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARYGFSGSRFYDTWGQGTLVTVSS [SEQ ID NO: 73]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:74 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-180 scFv (also referred to as “ET140-30scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:9 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:10, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 3. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:9, as shownin Table 3. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(H) comprising amino acids having the sequence setforth in SEQ ID NO:9, as shown in Table 3. In certain embodiments, theextracellular antigen-binding domain comprises a V_(L) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:10, asshown in Table 3. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:10, as shown in Table 3. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:9 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:10, as shown in Table 3. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:101 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:102 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:103 or conservative modifications thereof, as shownin Table 3. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:104 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:105 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:106 orconservative modifications thereof, as shown in Table 3. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:101 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:102 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:103 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:104 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:105 or conservative modifications thereof, and aV_(L) CDR3 comprising amino acids having the sequence set forth in SEQID NO:106 or conservative modifications thereof, as shown in Table 3. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:101, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:102, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:103, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:104, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:105, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:106.

TABLE 3 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GGTFSSYA IIPILGIAARSGYSKSIVSYMD [SEQ ID [SEQ ID [SEQ ID NO: 101] NO: 102] NO: 103] V_(L)SSNIGSNV RNN AAWDDSLSGYV [SEQ ID [SEQ ID [SEQ ID NO: 104] NO: 105]NO: 106] Full EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQ V_(H)GLEWMGRIIPILGIANYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARSGYSKSIVSYMDYWGQGTLVTVSS [SEQ ID NO: 9] DNAGaggtccagctggtgcagtctggagctgaggtgaagaagcctgggtcctcggtgaaggtctcctgcaaggcttctggaggcaccttcagcagctatgctatcagctgggtgcgacaggcccctggacaagggcttgagtggatgggaaggatcatccctatccttggtatagcaaactacgcacagaagttccagggcagagtcaccatgaccgaggacacatctacagacacagcctacatggagctgagcagcctgagatctgaggacacggccgtgtattactgtgcgcgctctggttactctaaatctatcgtttcttacatggattactggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 11] FullLPVLTQPPSTSGTPGQRVTVSCSGSSSNIGSNVVFWYQQLPGT V_(L)APKLVIYRNNQRPSGVPDRFSVSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGYVFGTGTKVTVLG [SEQ ID NO: 10] DNACtgcctgtgctgactcagcccccctccacgtctgggacccccgggcagagggtcaccgtctcttgttctggaagcagctccaacatcggaagtaatgttgtattctggtaccagcagctcccaggcacggcccccaaacttgtcatctataggaataatcaacggccctcaggggtccctgaccgattctctgtctccaagtctggcacctcagcctccctggccatcagtgggctccggtccgaggacgaggctgattattattgtgcagcttgggatgacagcctgagtggttatgtcttcggaactgggaccaaggtcaccgtcctaggt [SEQ ID NO: 12] scFvLPVLTQPPSTSGTPGQRVTVSCSGSSSNIGSNVVFWYQQLPGTAPKLVIYRNNQRPSGVPDRFSVSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCARSGYSKSIVSYMDYWGQGTLVTVSS [SEQ ID NO: 74]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:75 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-172 scFv (also referred to as “ET140-22scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:13 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:14, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 4. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:13, asshown in Table 4. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:13, as shown in Table 4. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:14, as shown in Table 4. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:14, as shown in Table 4. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:13 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:14, as shown in Table 4. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:107 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:108 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:109 or conservative modifications thereof, as shownin Table 4. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:110 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:111 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:112 orconservative modifications thereof, as shown in Table 4. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:107 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:108 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:109 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:110 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:111 or conservative modifications thereof, and aV_(L) CDR3 comprising amino acids having the sequence set forth in SEQID NO:112 or conservative modifications thereof, as shown in Table 4. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:107, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:108, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:109, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:110, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:111, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:112.

TABLE 4 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYTFTSYY INPSGGST ARSQWGGVLDY[SEQ ID [SEQ ID [SEQ ID NO: 107] NO: 108] NO: 109] V_(L) SSNIGARYD GNNQSYDSSLSASV [SEQ ID [SEQ ID [SEQ ID NO: 110] NO: 111] NO: 112] FullEVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQ V_(H)GLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSQWGGVLDYWGQGTLVTVSS [SEQ ID NO: 13] DNAGaggtccagctggtacagtctggggctgaggtgaagaagcctggggcctcagtgaaggtttcctgcaaggcatctggatacaccttcaccagctactatatgcactgggtgcgacaggcccctggacaagggcttgagtggatgggaataatcaaccctagtggtggtagcacaagctacgcacagaagttccagggcagagtcaccatgaccagggacacgtccacgagcacagtctacatggagctgagcagcctgagatctgaggacacggccgtgtattactgtgcgcgctctcagtggggtggtgttctggattactggggtcaaggtactctggtgac cgtctcctca [SEQ ID NO: 15]Full QSVVTQPPSVSGAPGQRVTISCSGSSSNIGARYDVQWYQQLPG V_(L)TAPKLLIFGNNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSASVFGGGTKLTVLG [SEQ ID NO: 14] DNACagtctgtcgtgacgcagccgccctcagtgtctggggccccagggcagagggtcaccatctcctgcagtgggagcagctccaacatcggggcacgttatgatgttcagtggtaccagcagcttccaggaacagcccccaaactcctcatctttggtaacaacaatcggccctcaggggtccctgaccgattctctggctccaagtctggcacgtcagcctccctggccatcactgggctccaggctgaggatgaggctgattattactgccagtcctatgacagcagcctgagtgcttcggtgttcggcggagggaccaagctgaccgtcctaggt [SEQ ID NO: 16] scFvQSVVTQPPSVSGAPGQRVTISCSGSSSNIGARYDVQWYQQLPGTAPKLLIFGNNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSASVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSQWGGVLDYWGQGTLVTVSS [SEQ ID NO: 75]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:76 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-157 scFv (also referred to as “ET140-7scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:17 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:18, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 5. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:17, asshown in Table 5. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:17, as shown in Table 5. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:18, as shown in Table 5. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:18, as shown in Table 5. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:17 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:18, as shown in Table 5. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:113 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:114 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:115 or conservative modifications thereof, as shownin Table 5. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:116 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:117 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:118 orconservative modifications thereof, as shown in Table 5. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:113 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:114 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:115 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:116 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:117 or conservative modifications thereof, and aV_(L) CDR3 comprising amino acids having the sequence set forth in SEQID NO:118 or conservative modifications thereof, as shown in Table 5. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:113, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:114, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:115, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:116, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:117, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:118.

TABLE 5 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GGTFSSYA IIPILGIAARTGYESWGSYEVIDR [SEQ ID [SEQ ID [SEQ ID NO: 113] NO: 114] NO: 115]V_(L) SSNIGSNT SNN AAWDDSLNGVV [SEQ ID [SEQ ID [SEQ ID NO: 116] NO: 117]NO: 118] Full QVQLVESGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQ V_(H)GLEWMGRIIPILGIANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARTGYESWGSYEVIDRWGQGTLVTVSS [SEQ ID NO: 17] DNACaggtgcagctggtggagtctggggctgaggtgaagaagcctgggtcctcggtgaaggtctcctgcaaggcttctggaggcaccttcagcagctatgctatcagctgggtgcgacaggcccctggacaagggcttgagtggatgggaaggatcatccctatccttggtatagcaaactacgcacagaagttccagggcagagtcacgattaccgcggacgaatccacgagcacagcctacatggagctgagcagcctgagatctgaggacacggccgtatattactgtgcgcgcactggttacgaatcttggggttcttacgaagttatcgatcgttggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 19] FullQAVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYRQLPGT V_(L)APKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGVVFGGGTKLTVLG [SEQ ID NO: 18] DNACaggctgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggaagcagctccaacatcggaagtaatactgtaaactggtaccggcagctcccaggaacggcccccaaactcctcatctatagtaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtgggctccagtctgaggatgaggctgattattactgtgcagcatgggatgacagcctgaatggtgtggtattcggcggagggaccaagctgaccgtcctaggt [SEQ ID NO: 20] scFvQAVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYRQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVESGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARTGYESWGSYEVIDRWGQGTLVTVSS [SEQ ID NO: 76]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:77 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-153 scFv (also referred to as “ET140-3scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:21 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:22, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 6. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:21, asshown in Table 6. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:21, as shown in Table 6. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:22, as shown in Table 6. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:22, as shown in Table 6. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:21 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:22, as shown in Table 6. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:119 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:120 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:121 or conservative modifications thereof, as shownin Table 6. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:122 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:123 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:124 orconservative modifications thereof, as shown in Table 6. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:119 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:120 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:121 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:122 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:123 or conservative modifications thereof, and aV_(L) CDR3 comprising amino acids having the sequence set forth in SEQID NO:124 or conservative modifications thereof, as shown in Table 6. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:119, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:120, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:121, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:122, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:123, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:124.

TABLE 6 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GGTFSSYA IIPILGIAARGGYYSHDMWS [SEQ ID [SEQ ID [SEQ ID NO: 119] NO: 120] NO: 121] V_(L)SSNIGSNS SNN ATWDDNLNVHYV [SEQ ID [SEQ ID [SEQ ID NO: 122] NO: 123]NO: 124] Full QVQLVESGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQ V_(H)GLEWMGRIIPILGIANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARTGYESWGSYEVIDRWGQGTLVTVSS [SEQ ID NO: 21] DNACaggtgcagctggtgcagtctggggctgaggtgaagaagcctgggtcctcggtgaaggtctcctgcaaggcttctggaggcaccttcagcagctatgctatcagctgggtgcgacaggcccctggacaagggcttgagtggatgggaaggatcatccctatccttggtatagcaaactacgcacagaagttccagggcagagtcacgattaccgcggacaaatccacgagcacagcctacatggagctgagcagcctgagatctgaggacacggccgtgtattactgtgcgcgcggtggttactactctcatgacatgtggtctgaagattggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 23] FullLPVLTQPPSASGTPGQRVTISCSGRSSNIGSNSVNWYRQLPGA V_(L)APKLLIYSNNQRPPGVPVRFSGSKSGTSASLAISGLQSEDEATYYCATWDDNLNVHYVFGTGTKVTVLG [SEQ ID NO: 22] DNACtgcctgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggacgcagttccaacatcgggagtaattctgttaactggtatcgacaactcccaggagcggcccccaaactcctcatctatagtaataatcagcggcccccaggggtccctgtgcgattctctggctccaagtctggcacctcagcctccctggccatcagtgggctccagtctgaagatgaggccacttattactgtgcaacatgggatgacaatctgaatgttcactatgtcttcggaactgggaccaaggtcaccgtcctaggt [SEQ ID NO: 24] scFvLPVLTQPPSASGTPGQRVTISCSGRSSNIGSNSVNWYRQLPGAAPKLLIYSNNQRPPGVPVRFSGSKSGTSASLAISGLQSEDEATYYCATWDDNLNVHYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGGYYSHDMWSEDWGQGTLVTVSS [SEQ ID NO: 77]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:78 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-201 scFv (also referred to as “ET140-51scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:25 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:26, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 7. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:25, asshown in Table 7. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:25, as shown in Table 7. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:26, as shown in Table 7. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:26, as shown in Table 7. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:25 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:26, as shown in Table 7. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:125 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:126 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:127 or conservative modifications thereof, as shownin Table 7. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:128 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:129 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:130 orconservative modifications thereof, as shown in Table 7. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:125 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:126 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:127 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:128 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:129 or conservative modifications thereof, and aV_(L) CDR3 comprising amino acids having the sequence set forth in SEQID NO:130 or conservative modifications thereof, as shown in Table 7. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:125, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:126, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:127, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:128, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:129, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:130.

TABLE 7 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GGSISNSNW IYHSGST ARRDNWKTP[SEQ ID [SEQ ID TTKIDGFDI NO: 125] NO: 126] [SEQ ID NO: 127] V_(L)SGYSNYK VGTGGIVG GADHGSG [SEQ ID [SEQ ID SNFVYV NO: 128] NO: 129] SEQ IDNO: 130] Full QVQLQESGPGLVKPSGTLSLTCGVSGGSISNSNWWSWVRQPPG V_(H)KGLEWIGEIYHSGSTKYNPSLRSRVTISVDKSKNQFSLKLSSVTAADTAVYYCARRDNWKTPTTKIDGFDIWGQGTMVTVSS [SEQ ID NO: 25] DNACaggtgcagctgcaggagtcgggcccaggactggtgaagccttcggggaccctgtccctcacctgcggtgtctctggtggctccatcagcaatagtaactggtggagttgggtccgccagccccccgggaaggggctggagtggattggggaaatctatcatagtgggagcaccaagtacaacccgtccctcaggagtcgagtcaccatatcagtagacaagtccaagaaccagttctccctaaaattgagctctgtgaccgccgcggacacggccgtatattactgtgcgagacgagataactggaagacccccactaccaaaattgatggttttgatatctggggccaagggacaatggtcaccgtctcttca [SEQ ID NO: 27] FullQPVLTQPPSASASLGASVTLTCTLSSGYSNYKVDWYQQRPGKG V_(L)PRFVMRVGTGGIVGSKGDGIPDRFSVLGSGLNRYLTIKNIQEEDEGDYHCGADHGSGSNFVYVFGTGTKVTVLG [SEQ ID NO: 26] DNACagcctgtgctgactcagccaccttctgcatcagcctccctgggagcctcggtcacactcacctgcaccctgagcagcggctacagtaattataaagtggactggtaccagcagagaccagggaagggcccccggtttgtgatgcgagtgggcactggtgggattgtgggatccaagggggatggcatccctgatcgcttctcagtcttgggctcaggcctgaatcggtacctgaccatcaagaacatccaggaagaagatgagggtgactatcactgtggggcagaccatggcagtgggagcaacttcgtgtatgtcttcggaactgggaccaaggtcaccgt cctaggt [SEQ ID NO: 28] scFvQPVLTQPPSASASLGASVTLTCTLSSGYSNYKVDWYQQRPGKGPRFVMRVGTGGIVGSKGDGIPDRFSVLGSGLNRYLTIKNIQEEDEGDYHCGADHGSGSNFVYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLQESGPGLVKPSGTLSLTCGVSGGSISNSNWWSWVRQPPGKGLEWIGEIYHSGSTKYNPSLRSRVTISVDKSKNQFSLKLSSVTAADTAVYYCARRDNWKTPTTKIDGFDIWGQGTM VTVSS [SEQ ID NO: 78]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:79 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-167 scFv (also referred to as “ET140-17scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:29 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:30, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 8. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:29, asshown in Table 8. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:29, as shown in Table 8. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:30, as shown in Table 8. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:30, as shown in Table 8. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:29 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:30, as shown in Table 8. In certain embodiments, the anti-BCMAcomprises a V_(H) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO:131 or conservative modifications thereof, a V_(H)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:132 or conservative modifications thereof, and a V_(H) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:133 orconservative modifications thereof, as shown in Table 8. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:134 or conservative modifications thereof, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:135 orconservative modifications thereof, and a V_(L) CDR3 comprising aminoacids having the sequence set forth in SEQ ID NO:136 or conservativemodifications thereof, as shown in Table 8. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) CDR1 comprisingamino acids having the sequence set forth in SEQ ID NO:131 orconservative modifications thereof, a V_(H) CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO:132 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:133 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO:134 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:135 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:136 orconservative modifications thereof, as shown in Table 8. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:131, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:132, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:133, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:134, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:135, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:136.

TABLE 8 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYTFTGYY INPNSGGT ARSQWGSSWDY[SEQ ID [SEQ ID [SEQ ID NO: 131] NO: 132] NO: 133] V_(L) QSISSY AASQQSYSTPPT [SEQ ID [SEQ ID [SEQ ID NO: 134] NO: 135] NO: 136] FullQVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQ V_(H)GLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARSQWGSSWDYWGQGTLVTVSS [SEQ ID NO: 29] DNACaggtccagctggtacagtctggggctgaggtgaagaagcctggggcctcagtgaaggtctcctgcaaggcttctggatacaccttcaccggctactatatgcactgggtgcgacaggcccctggacaagggcttgagtggatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcagggcagggtcaccatgaccagggacacgtccatcagcacagcctacatggagctgagcaggctgagatctgacgacacggccgtgtattactgtgcgcgctctcagtggggttcttcttgggattactggggtcaaggtactctggtgac cgtctcctca [SEQ ID NO: 31]Full DIQLTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKA V_(L)PKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIKR [SEQ ID NO: 30] DNAGacatccagttgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgccgggcaagtcagagcattagcagctatttaaattggtatcagcagaaaccagggaaagcccctaagctcctgatctatgctgcatccagtttgcaaagtggggtcccatcaaggttcagtggcagtggatctgggacagatttcactctcaccatcagcagtctgcaacctgaagattttgcaacttactactgtcaacagagttacagtacccctccgacgttcggccaagggaccaaggtggagatcaaacgt [SEQ ID NO: 32] scFvDIQLTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARSQWGSSWDYWGQGTLVTVSS [SEQ ID NO: 79]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:80 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-163 scFv (also referred to as “ET140-13scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:33 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:34, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 9. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:33, asshown in Table 9. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:33, as shown in Table 9. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:34, as shown in Table 9. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:34, as shown in Table 9. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:33 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:34, as shown in Table 9. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:137 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:138 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:139 or conservative modifications thereof, as shownin Table 9. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:140 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:141 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:142 orconservative modifications thereof, as shown in Table 9. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:137 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:138 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:139 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:140 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:141 or conservative modifications thereof, and aV_(L) CDR3 comprising amino acids having the sequence set forth in SEQID NO:142 or conservative modifications thereof, as shown in Table 9. Incertain embodiments, the extracellular antigen-binding domain comprisesa V_(H) CDR1 comprising amino acids having the sequence set forth in SEQID NO:137, a V_(H) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:138, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO:139, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:140, a V_(L) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:141, and a V_(L)CDR3 comprising amino acids having the sequence set forth in SEQ IDNO:142.

TABLE 9 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYTFTGYY INPNSGGTARSSYHLYGYDS [SEQ ID [SEQ ID [SEQ ID NO: 137] NO: 138] NO: 139] V_(L)NDYTNYK VGPGGIVG GADHGTGSNFVYV [SEQ ID [SEQ ID [SEQ ID NO: 140] NO: 141]NO: 142] Full EVQLVESGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQ V_(H)GLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARSSYHLYGYDSWGQGTLVTVSS [SEQ ID NO: 33] DNAGaggtgcagctggtggagtccggggctgaggtgaagaagcctggggcctcagtgaaggtctcctgcaaggcttctggatacaccttcaccggctactatatgcactgggtgcgacaggcccctggacaagggcttgagtggatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcagggcagggtcaccatgaccagggacacgtccatcagcacagcctacatggagctgagcaggctgagatctgacgacacggccgtatattactgtgcgcgctcttcttaccatctgtacggttacgattcttggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 35] FullQPVLTQPPSASASLGASVTLTCTLSNDYTNYKVDWYQQRPGKG V_(L)PRFVMRVGPGGIVGSKGDGIPDRFSVLGSGLNRYLTIKNIQEEDESDYHCGADHGTGSNFVYVFGGGTKLTVLG [SEQ ID NO: 34] DNACagcctgtgctgactcagccaccttctgcatcagcctccctgggagcctcggtcactctcacctgcaccctgagcaacgactacactaattataaagtggactggtaccagcagagaccagggaagggcccccggtttgtgatgcgagtgggccctggtgggattgtgggatccaagggggatggcatccctgatcgcttctcagtcttgggctcaggcctgaatcgatacctgaccatcaagaacatccaggaggaggatgagagtgactaccactgtggggcggaccatggcaccgggagcaacttcgtgtacgtgttcggcggagggaccaagctgaccgt cctaggt [SEQ ID NO: 36] scFvQPVLTQPPSASASLGASVTLTCTLSNDYTNYKVDWYQQRPGKGPRFVMRVGPGGIVGSKGDGIPDRFSVLGSGLNRYLTIKNIQEEDESDYHCGADHGTGSNFVYVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVESGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARSSYHLYGYDSWGQGTLVTVSS [SEQ ID NO: 80]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:81 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-207 scFv (also referred to as “ET140-57scFv”).

In certain embodiment, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:37 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:38, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:98. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 10. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:37, asshown in Table 10. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:37, as shown in Table 10. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:38, as shown in Table 10. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:38, as shown in Table 10. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:37 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:38, as shown in Table 10. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:143 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:144 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:145 or conservative modifications thereof, as shownin Table 10. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO:146 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:147 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:148 orconservative modifications thereof, as shown in Table 10. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ IDNO:143 or conservative modifications thereof, a V_(H) CDR2 comprisingamino acids having the sequence set forth in SEQ ID NO:144 orconservative modifications thereof, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:145 or conservativemodifications thereof, a V_(L) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:146 or conservative modificationsthereof, a V_(L) CDR2 comprising amino acids having the sequence setforth in SEQ ID NO:147 or conservative modifications thereof, and aV_(L) CDR3 comprising amino acids having the sequence set forth in SEQID NO:148 or conservative modifications thereof, as shown in Table 10.In certain embodiments, the extracellular antigen-binding domaincomprises a V_(H) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO:143, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:144, a V_(H) CDR3 comprising amino acidshaving the sequence set forth in SEQ ID NO:145, a V_(L) CDR1 comprisingamino acids having the sequence set forth in SEQ ID NO:146, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO:147,and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO:148.

TABLE 10 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GGTFSSYA IIPIFSTAARQPWTWYSPYDQ [SEQ ID [SEQ ID [SEQ ID NO: 143] NO: 144] NO: 145] V_(L)SGYSNYK VDTGGIVG GADHGSGSNFVWV [SEQ ID [SEQ ID [SEQ ID NO: 146] NO: 147]NO: 148] Full QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQ V_(H)GLEWMGGIIPIFSTANYAQKFQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARQPWTWYSPYDQWGQGTLVTVSS [SEQ ID NO: 37] DNACaggtgcagctggtgcagtctggggctgaggtgaagaagcctgggtcctcggtgaaggtctcctgcaaggcttctggaggcaccttcagcagctatgctatcagctgggtgcgacaggcccctggacaagggcttgagtggatgggagggatcatccctatctttagtacagcaaactacgcacagaagttccagggcagagtcaccatgaccacagacacatccacgagcacagcctacatggagctgaggagcctgagatctgacgacacggccgtgtattactgtgcgcgccagccgtggacttggtactctccgtacgatcagtggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 39] FullQPVLTQPSSASASLGASVTLTCTLSSGYSNYKVDWYQQRPGKG V_(L)PRFLMRVDTGGIVGSKGDGIPDRFSVSGSGLNRYLTIKNIQEEDESDYHCGADHGSGSNFVWVFGGGTKLTVLG [SEQ ID NO: 38] DNACagcctgtgctgactcagccaccttctgcatcagcctccctgggagcctcggtcacctcacctgcaccctgagcagcggctacagtaattataaagtggactggtatcaacagagaccagggaagggcccccggtttctgatgcgagtagacaccggtgggattgtgggatccaagggggatggcatccctgatcgcttctcagtctcgggctcaggtctgaatcggtacctgaccatcaagaacattcaggaagaggatgagagtgactaccactgtggggcagaccatggcagtgggagcaacttcgtgtgggtgttcggcggagggaccaagctgaccgtc ctaggt [SEQ ID NO: 40] scFvQPVLTQPPSASASLGASVTLTCTLSSGYSNYKVDWYQQRPGKGPRFLMRVDTGGIVGSKGDGIPDRFSVSGSGLNRYLTIKNIQEEDESDYHCGADHGSGSNFVWVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFSTANYAQKFQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARQPWTWYSPYDQWGQGTLVTVSS [SEQ ID NO: 81]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:82 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-165 scFv (also referred to as “ET140-15scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:41 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:42, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 11. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:41, asshown in Table 11. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:41, as shown in Table 11. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:42, as shown in Table 11. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:42, as shown in Table 11. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:41 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:42, as shown in Table 11. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:149 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 150 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 151 or conservative modifications thereof, as shownin Table 11. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 152 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO: 153 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 154or conservative modifications thereof, as shown in Table 11. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:147 or conservative modifications thereof, a V_(H) CDR2 comprising aminoacids having the sequence set forth in SEQ ID NO: 150 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 151 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 152 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ ID NO:153 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 154 orconservative modifications thereof, as shown in Table 11. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:149, a V_(H) CDR2 comprising amino acids having the sequence set forthin SEQ ID NO: 150, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 151, a V_(L) CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 152, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 153,and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO: 154.

TABLE 11 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GFTFSTYA ITPGGDRT ARYYGYMIDM[SEQ ID [SEQ ID [SEQ ID NO: 149] NO: 150] NO: 151] V_(L) QSLLHSNGYNY LGSMQALQTPLT [SEQ ID [SEQ ID [SEQ ID NO: 152] NO: 153] NO: 154] FullEVQLVETGGGLVQPGGSLRLSCAASGFTFSTYAMTWVRQAPGK V_(H)GLEWVSAITPGGDRTYYADSVKGRFTISRDNSRNTLYLQMNSLRAEDTAVYYCARYYGYMIDMWGQGTLVTVSS [SEQ ID NO: 41] DNAGaggtgcagctggtggagactgggggaggcctggtacagcctggggggtccctgagactctcctgtgctgcctctggattcacctttagcacctatgccatgacctgggtccgccaggctccagggaaggggctggagtgggtctcagctattactcctggtggtgatcgcacatactacgcagactccgtgaagggccgtttcactatctccagagacaattccaggaacacgctgtatctgcaaatgaacagcctgagagccgaggacacggccgtatattactgtgcgcgctactacggttacatgatcgatatgtggggtcaaggtactctggtgaccgt ctcctca [SEQ ID NO: 43] FullDVVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQ V_(L)KPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAE DVGVYYCMQALQTPLTFGGGTKVEIKR[SEQ ID NO: 42] DNA Gatgttgtgatgactcagtctccactctccctgcccgtcacccctggagagccggcctccatctcctgcaggtctagtcagagcctcctgcatagtaatggatacaactatttggattggtacctgcagaagccagggcagtctccacagctcctgatctatttgggttctaatcgggcctccggggtccctgacaggttcagtggcagtggatcaggcacagattttacactgaaaatcagcagagtggaggctgaggatgttggggtttattactgcatgcaagctctacaaactcctctcactttcggcggagggaccaaggtggaaatcaaacgt [SEQ ID NO: 44] scFvDVVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPLTFGGGTKVEIKRSRGGGGSGGGGSGGGGSLEMAEVQLVETGGGLVQPGGSLRLSCAASGFTFSTYAMTWVRQAPGKGLEWVSAITPGGDRTYYADSVKGRFTISRDNSRNTLYLQMNSLRAEDTAVYYCARYYGYMIDMWGQGTLVTVSS [SEQ ID NO: 82]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:83 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-188 scFv (also referred to as “ET140-38scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:45 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:46, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 12. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:45, asshown in Table 12. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:45, as shown in Table 12. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:46, as shown in Table 12. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:46, as shown in Table 12. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:45 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:46, as shown in Table 12. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:155 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 156 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 157 or conservative modifications thereof, as shownin Table 12. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 158 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO: 159 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 160or conservative modifications thereof, as shown in Table 12. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:155 or conservative modifications thereof, a V_(H) CDR2 comprising aminoacids having the sequence set forth in SEQ ID NO: 156 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 157 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 158 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ ID NO:159 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 160 orconservative modifications thereof, as shown in Table 12. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:155, a V_(H) CDR2 comprising amino acids having the sequence set forthin SEQ ID NO: 156, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 157, a V_(L) CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 158, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 159,and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO: 160.

TABLE 12 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYTFTGYY INPNSGGT ARSQWGGTYDY[SEQ ID [SEQ ID [SEQ ID NO: 155] NO: 156] NO: 157] V_(L) SSNIGSNT SNNAAWDDSLNGWV [SEQ ID [SEQ ID [SEQ ID NO: 158] NO: 159] NO: 160] FullQMQLVQSGAEVKKPGASVKVSCKASGYTFTGYYVHWLRQAPGQ V_(H)GLEWMGWINPNSGGTNNAQEFQGRITMTRDTSINTAYMELSRLRSDDTAVYYCARSQWGGTYDYWGQGTLVTVSS [SEQ ID NO: 45] DNACagatgcagctggtgcagtctggggctgaggtgaagaagcctggggcctcagtgaaggtctcctgcaaggcttctggatacaccttcaccggctattatgtacactggttgcgacaggcccctggacaagggcttgagtggatgggttggatcaaccctaacagtggcggcacaaacaatgcacaggagtttcaaggcaggatcaccatgaccagggacacgtccatcaacacagcctacatggagctgagcaggctgagatctgacgacacggccgtgtattactgtgcgcgctctcagtggggtggtacttacgattactggggtcaaggtactctggtgac cgtctcctca [SEQ ID NO: 47]Full SYVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQVPGT V_(L)APKLLIYSNNQRPSGVPDRFSGSKSGASASLAISWLQSEDEADYYCAAWDDSLNGWVFGGGTKLTVLG [SEQ ID NO: 46] DNATcctatgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggaagcagctccaacatcggaagtaatactgtaaactggtaccagcaggtcccaggaacggcccccaaactcctcatctatagtaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcgcctcagcctccctggccatcagttggctccagtctgaggatgaggctgattattactgtgcagcatgggatgacagcctgaatggttgggtgttcggcggagggaccaagctgaccgtcctaggt [SEQ ID NO: 48] scFvSYVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQVPGTAPKLLIYSNNQRPSGVPDRFSGSKSGASASLAISWLQSEDEADYYCAAWDDSLNGWVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAQMQLVQSGAEVKKPGASVKVSCKASGYTFTGYYVHWLRQAPGQGLEWMGWINPNSGGTNNAQEFQGRITMTRDTSINTAYMELSRLRSDDTAVYYCARSQWGGTYDYWGQGTLVTVSS [SEQ ID NO: 83]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:84 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-196 scFv (also referred to as “ET140-46scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:49 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:50, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 13. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:49, asshown in Table 13. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:49, as shown in Table 13. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:50, as shown in Table 13. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:50, as shown in Table 13. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:49 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:50, as shown in Table 13. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:161 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 162 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 163 or conservative modifications thereof, as shownin Table 13. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 164 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:165 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:166 orconservative modifications thereof, as shown in Table 13. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:161 or conservative modifications thereof, a V_(H) CDR2 comprising aminoacids having the sequence set forth in SEQ ID NO: 162 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 163 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 164 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:165 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:166 orconservative modifications thereof, as shown in Table 13. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:161, a V_(H) CDR2 comprising amino acids having the sequence set forthin SEQ ID NO: 162, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 163, a V_(L) CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 164, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO:165,and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO:166.

TABLE 13 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYDFTTYW IYPGDSDT ARMWTFSQDG[SEQ ID [SEQ ID [SEQ ID NO: 161] NO: 162] NO: 163] V_(L) SSNIGSYT SNNAAWDDSLNGYV [SEQ ID [SEQ ID [SEQ ID NO: 164] NO: 165] NO: 166] FullEVQLVQSGAEVKKPGESLKISCKGSGYDFTTYWIGWVRQMPGK V_(H)GLEWMGIIYPGDSDTRYSPSVRGRVTISADKSINTAYLQWSSLEASDTAMYYCARMWTFSQDGWGQGTLVTVSS [SEQ ID NO: 49] DNAgaggtgcagctggtgcagtctggagcagaggtgaaaaagccgggggagtctctgaagatctcctgtaagggttctggatatgactttaccacctactggatcgggtgggtgcgccagatgcccgggaagggcctggagtggatggggatcatctatcctggtgactctgataccagatacagcccgtccgtccgaggccgggtcaccatctcagccgacaagtccatcaacaccgcctatttgcagtggagtagcctggaggcctccgacaccgccatgtattactgtgcgcgcatgtggactttctctcaggatggttggggtcaaggtactctggtgaccgt ctcctca [SEQ ID NO: 51] FullQAVLTQPPSASGTPGQRVTISCSGSSSNIGSYTVSWYQQLPGT V_(L)APKFLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGYVFGTGTKVTVLG [SEQ ID NO: 50] DNACaggctgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggaagcagctccaacatcggaagttatactgtaagctggtaccagcaactcccaggaacggcccccaaattcctcatctattctaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtgggctccagtctgaggatgaggctgattattactgtgctgcatgggatgacagcctgaatggttatgtcttcggaactgggaccaaggtcaccgtcctaggt [SEQ ID NO: 52] scFvQAVLTQPPSASGTPGQRVTISCSGSSSNIGSYTVSWYQQLPGTAPKFLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYDFTTYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSVRGRVTISADKSINTAYLQWSSLEASDTAMYYCARMWTFSQDGWGQGTLVTVSS [SEQ ID NO: 84]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:85 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-204 scFv (also referred to as “ET140-54scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:53 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:54, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 14. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:53, asshown in Table 14. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:53, as shown in Table 14. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:54, as shown in Table 14. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:54, as shown in Table 14. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:53 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:54, as shown in Table 14. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:167 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:168 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:169 or conservative modifications thereof, as shownin Table 14. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 170 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO: 171 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 172or conservative modifications thereof, as shown in Table 14. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:167 or conservative modifications thereof, a V_(H) CDR2 comprising aminoacids having the sequence set forth in SEQ ID NO: 168 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 169 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 170 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ ID NO:171 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 172 orconservative modifications thereof, as shown in Table 14. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:167, a V_(H) CDR2 comprising amino acids having the sequence set forthin SEQ ID NO: 168, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 169, a V_(L) CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 170, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 171,and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO: 172.

TABLE 14 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYTFIDYY INPNSGGT ARSQRDGYMDY[SEQ ID [SEQ ID [SEQ ID NO: 167] NO: 168] NO: 169] V_(L) ISCTGTSSD EDSSSNTRSSTLV [SEQ ID [SEQ ID [SEQ ID NO: 170] NO: 171] NO: 172] FullEVQLVQSGAEMKKPGASLKLSCKASGYTFIDYYVYWMRQAPGQ V_(H)GLESMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAMYYCARSQRDGYMDYWGQGTLVTVSS [SEQ ID NO: 53] DNAGaagtgcagctggtgcagtctggggctgagatgaagaagcctggggcctcactgaagctctcctgcaaggcttctggatacaccttcatcgactactatgtatactggatgcgacaggcccctggacaagggcttgagtccatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcagggcagggtcaccatgaccagggacacgtccatcagcacagcctacatggagctgagcaggctgagatctgacgacaccgccatgtattactgtgcgcgctcccagcgtgacggttacatggattactggggtcaaggtactctggtgac cgtctcctca [SEQ ID NO: 55]Full QSALTQPASVSASPGQSIAISCTGTSSDVGWYQQHPGKAPKLM V_(L)IYEDSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSNTRSSTLVFGGGTKLTVLG [SEQ ID NO: 54] DNACaatctgccctgactcagcctgcctccgtgtctgcgtctcctggacagtcgatcgccatctcctgcactggaaccagcagtgacgttggttggtatcaacagcacccaggcaaagcccccaaactcatgatttatgaggacagtaagcggccctcaggggtttctaatcgcttctctggctccaagtctggcaacacggcctccctgaccatctctgggctccaggctgaggacgaggctgattattactgcagctcaaatacaagaagcagcactttggtgttcggcggagggaccaagctgaccgtcctaggt [SEQ ID NO: 56] scFvQSALTQPASVSASPGQSIAISCTGTSSDVGWYQQHPGKAPKLMIYEDSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSNTRSSTLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEMKKPGASLKLSCKASGYTFIDYYVYWMRQAPGQGLESMGWINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAMYYCARSQRDGYMDYWGQGTLVTVSS [SEQ ID NO: 85]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:86 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-190 scFv (also referred to as “ET140-40scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:57 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:58, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 15. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:57, asshown in Table 15. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:57, as shown in Table 15. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:58, as shown in Table 15. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:58, as shown in Table 15. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:57 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:58, as shown in Table 15. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:173 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 174 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 175 or conservative modifications thereof, as shownin Table 15. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 176 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO: 177 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 178or conservative modifications thereof, as shown in Table 15. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:173 or conservative modifications thereof, a V_(H) CDR2 comprising aminoacids having the sequence set forth in SEQ ID NO: 174 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 175 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 176 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ ID NO:177 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 178 orconservative modifications thereof, as shown in Table 15. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:173, a V_(H) CDR2 comprising amino acids having the sequence set forthin SEQ ID NO: 174, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 175, a V_(L) CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 176, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 177,and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO: 178.

TABLE 15 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYTFTDYY INPNSGGT ARSPYSGVLDK[SEQ ID [SEQ ID [SEQ ID NO: 173] NO: 174 NO: 175] V_(L) SSNIGAGFD GNSQSYDSSLSGYV [SEQ ID [SEQ ID [SEQ ID NO: 176] NO: 177] NO: 178] FullQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMHWVRQAPGQ V_(H)RLEWMGWINPNSGGTNYAQKFQDRITVTRDTSSNTGYMELTRLRSDDTAVYYCARSPYSGVLDKWGQGTLVTVSS [SEQ ID NO: 57] DNACaggtccagctggtacagtctggggctgaggtgaagaagcctggggcctcagtgaaggtctcctgcaaggcttctggatacaccttcaccgactactatatgcactgggtgcgacaggcccctggacaacggcttgagtggatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcaggacaggatcaccgtgaccagggacacctccagcaacacaggctacatggagctgaccaggctgagatctgacgacacggccgtgtattactgtgcgcgctctccgtactctggtgttctggataaatggggtcaaggtactctggtgac cgtctcctca [SEQ ID NO: 59]Full QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGFDVHWYQQLPG V_(L)TAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVLG [SEQ ID NO: 58] DNACagtctgtgctgacgcagccgccctcagtgtctggggccccagggcagagggtcaccatctcctgcactgggagcagctccaacatcggggcaggttttgatgtacactggtaccagcagcttccaggaacagcccccaaactcctcatctatggtaacagcaatcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcactgggctccaggctgaggatgaggctgattattactgccagtcctatgacagcagcctgagtggttatgtcttcggaactgggaccaaggtcaccgtcctaggt [SEQ ID NO: 60] scFvQSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGFDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMHWVRQAPGQRLEWMGWINPNSGGTNYAQKFQDRITVTRDTSSNTGYMELTRLRSDDTAVYYCARSPYSGVLDKWGQGTLVTVSS [SEQ ID NO: 86]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO: 87 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-187 scFv (also referred to as “ET140-37scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:61 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:62, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 16. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:61, asshown in Table 16. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:61, as shown in Table 16. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:62, as shown in Table 16. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:62, as shown in Table 16. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:61 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:62, as shown in Table 16. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:179 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 180 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 181 or conservative modifications thereof, as shownin Table 16. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 182 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO: 183 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 184or conservative modifications thereof, as shown in Table 16. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:179 or conservative modifications thereof, a V_(H) CDR2 comprising aminoacids having the sequence set forth in SEQ ID NO: 180 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 181 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 182 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ ID NO:183 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 184 orconservative modifications thereof, as shown in Table 16. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:179, a V_(H) CDR2 comprising amino acids having the sequence set forthin SEQ ID NO: 180, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 181, a V_(L) CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 182, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 183and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO: 184.

TABLE 16 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GGTFSSYA IIPILGTAARSGYGSYRWEDS [SEQ ID [SEQ ID [SEQ ID NO: 179] NO: 180] NO: 181] V_(L)SSNIGSNY SNN AAWDDSLSASYV [SEQ ID [SEQ ID [SEQ ID NO: 182] NO: 183]NO: 184] Full QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQ V_(H)GLEWMGRIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSGYGSYRWEDSWGQGTLVTVSS [SEQ ID NO: 61] DNACaggtgcagctggtgcagtctggggctgaggtgaagaagcctgggtcctcggtgaaggtctcctgcaaggcttctggaggcaccttcagcagctatgctatcagctgggtgcgacaggcccctggacaagggcttgagtggatgggaaggatcatccctatccttggtacagcaaactacgcacagaagttccagggcagagtcacgattaccgcggacgaatccacgagcacagcctacatggagctgagcagcctgagatctgaggacacggccgtgtattactgtgcgcgctctggttacggttcttaccgttgggaagattcttggggtcaaggtactctggtgaccgtctcctca [SEQ ID NO: 63] FullQAVLTQPPSASGTPGQRVTISCSGSSSNIGSNYVFWYQQLPGT V_(L)APKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSASYVFGTGTKVTVLG [SEQ ID NO: 62] DNACaggctgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggaagcagctccaacatcggaagtaattacgtattctggtaccagcagctcccaggaacggcccccaaactcctcatctatagtaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtgggctccggtccgaggatgaggctgattattactgtgcagcatgggatgacagcctgagtgcctcttatgttttcggaactgggaccaaggtcaccgtcctaggt [SEQ ID NO: 64] scFvQAVLTQPPSASGTPGQRVTISCSGSSSNIGSNYVFWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSASYVFGTGTKVTVLGSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSGYGSYRWEDSWGQGTLVTVSS [SEQ ID NO: 87]

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises the amino acid sequence of SEQ ID NO:88 andspecifically binds to a BCMA polypeptide (e.g., a BCMA polypeptidehaving the amino acid sequence SEQ ID NO:71, or fragments thereof),which is designated as ET140-174 scFv (also referred to as “ET140-24scFv”).

In certain embodiments, the extracellular antigen-binding domain is ahuman scFv, which comprises a heavy chain variable region comprisingamino acids having the sequence set forth in SEQ ID NO:65 and a lightchain variable region comprising amino acids having the sequence setforth in SEQ ID NO:66, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain is a humanscFv-Fc fusion protein or full length human IgG with V_(H) and V_(L)regions or CDRs selected from Table 17. In certain embodiments, theextracellular antigen-binding domain comprises a V_(H) comprising anamino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%homologous to the amino acid sequence set forth in SEQ ID NO:65, asshown in Table 17. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) comprising amino acids havingthe sequence set forth in SEQ ID NO:65, as shown in Table 17. In certainembodiments, the extracellular antigen-binding domain comprises a V_(L)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:66, as shown in Table 17. In certain embodiments, the extracellularantigen-binding domain comprises a V_(L) comprising amino acids havingthe sequence set forth in SEQ ID NO:66, as shown in Table 17. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)comprising amino acids having the sequence set forth in SEQ ID NO:65 anda V_(L) comprising amino acids having the sequence set forth in SEQ IDNO:66, as shown in Table 17. In certain embodiments, the extracellularantigen-binding domain comprises a V_(H) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:185 or conservativemodifications thereof, a V_(H) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO: 186 or conservative modificationsthereof, and a V_(H) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO: 187 or conservative modifications thereof, as shownin Table 17. In certain embodiments, the extracellular antigen-bindingdomain comprises a V_(L) CDR1 comprising amino acids having the sequenceset forth in SEQ ID NO: 188 or conservative modifications thereof, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO: 189 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO: 190or conservative modifications thereof, as shown in Table 17. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:185 or conservative modifications thereof, a V_(H) CDR2 comprising aminoacids having the sequence set forth in SEQ ID NO: 186 or conservativemodifications thereof, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 187 or conservative modificationsthereof, a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO: 188 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ ID NO:189 or conservative modifications thereof, and a V_(L) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO: 190 orconservative modifications thereof, as shown in Table 17. In certainembodiments, the extracellular antigen-binding domain comprises a V_(H)CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:185, a V_(H) CDR2 comprising amino acids having the sequence set forthin SEQ ID NO: 186, a V_(H) CDR3 comprising amino acids having thesequence set forth in SEQ ID NO: 187, a V_(L) CDR1 comprising aminoacids having the sequence set forth in SEQ ID NO: 188, a V_(L) CDR2comprising amino acids having the sequence set forth in SEQ ID NO: 189,and a V_(L) CDR3 comprising amino acids having the sequence set forth inSEQ ID NO: 190.

TABLE 17 Anti- A BCMA polypeptide having the amino acid gensequence of SEQ ID NO: 71 CDRs 1 2 3 V_(H) GYSFTSYW IYPGDSDT ARYSGSFDN[SEQ ID [SEQ ID [SEQ ID NO: 185] NO: 186] NO: 187] V_(L) SSNIGSHS TNNAAWDGSLNGLV [SEQ ID [SEQ ID [SEQ ID NO: 188] NO: 189] NO: 190] FullEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGK V_(H)GLEWMGIIYPGDSDTRYSPSFQGHVTISADKSISTAYLQWSSLKASDTAMYYCARYSGSFDNWGQGTLVTVSS [SEQ ID NO: 65] DNAGaggtgcagctggtgcagtctggagcagaggtgaaaaagcccggggagtctctgaagatctcctgtaagggttctggatacagctttaccagctactggatcggctgggtgcgccagatgcccgggaaaggcctggagtggatggggatcatctatcctggtgactctgataccagatacagcccgtccttccaaggccacgtcaccatctcagctgacaagtccatcagcactgcctacctgcagtggagcagcctgaaggcctcggacaccgccatgtattactgtgcgcgctactctggttctttcgataactggggtcaaggtactctggtgaccgtctc ctca [SEQ ID NO: 67] FullSYELTQPPSASGTPGQRVTMSCSGTSSNIGSHSVNWYQQLPGT V_(L)APKLLIYTNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDGSLNGLVFGGGTKLTVLG [SEQ ID NO: 66] DNATcctatgagctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatgtcttgttctggaaccagctccaacatcggaagtcactctgtaaactggtaccagcagctcccaggaacggcccccaaactcctcatctatactaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtggcctccagtctgaggatgaggctgattattactgtgcagcatgggatggcagcctgaatggtctggtattcggcggagggaccaagctgaccgtcctaggt [SEQ ID NO: 68] scFvSYELTQPPSASGTPGQRVTMSCSGTSSNIGSHSVNWYQQLPGTAPKLLIYTNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDGSLNGLVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGHVTISADKSISTAYLQWSSLKASDTAMYYCARYSGSFDNWGQGTLVTVSS [SEQ ID NO: 88]

An extracellular antigen-binding domain (e.g., scFv) comprising V_(H)and/or V_(L) regions having high (i.e., 80% or greater) homology to theV_(H) and V_(L) regions of the sequences set forth above, can beobtained by mutagenesis (e.g., site-directed or PCR-mediatedmutagenesis), followed by testing of the encoded altered scFv forretained function (i.e., the binding affinity) using the binding assaysdescribed herein. In certain embodiments, a V_(H) sequence having atleast 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identitycontains substitutions (e.g., conservative substitutions to generateconservative modifications of a sequence), insertions or deletionsrelative to the reference sequence, but an extracellular antigen-bindingdomain (e.g., scFv) comprising that sequence retains the ability to bindto a BCMA polypeptide. In certain embodiments, a V_(L) sequence havingat least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identitycontains substitutions (e.g., conservative substitutions), insertions ordeletions relative to the reference sequence, but an extracellularantigen-binding domain (e.g., scFv) comprising that sequence retains theability to bind to a BCMA polypeptide. In certain embodiments, a totalof about 1 to about 10 amino acids have been substituted, insertedand/or deleted in the disclosed sequences. For example, and not by wayof limitation, a V_(H) sequence or a V_(L) sequence, can have up toabout one, up to about two, up to about three, up to about four, up toabout five, up to about six, up to about seven, up to about eight, up toabout nine or up to about ten amino acid residues that are modifiedand/or substituted. Non-limiting examples of conservative modificationsare provided below, e.g., within Table 18.

As used herein, the term “conservative sequence modifications” refers toamino acid modifications that do not significantly affect or alter thebinding characteristics of the presently disclosed CAR (e.g., theextracellular antigen-binding domain) comprising the amino acidsequence. Such conservative modifications include amino acidsubstitutions, additions and deletions. Modifications can be introducedinto the human scFv of the presently disclosed subject matter bystandard techniques known in the art, such as site-directed mutagenesisand PCR-mediated mutagenesis. Amino acids can be classified into groupsaccording to their physicochemical properties such as charge andpolarity. Conservative amino acid substitutions are ones in which theamino acid residue is replaced with an amino acid within the same group.For example, amino acids can be classified by charge: positively-chargedamino acids include lysine, arginine, histidine, negatively-chargedamino acids include aspartic acid, glutamic acid, neutral charge aminoacids include alanine, asparagine, cysteine, glutamine, glycine,isoleucine, leucine, methionine, phenylalanine, proline, serine,threonine, tryptophan, tyrosine, and valine. In addition, amino acidscan be classified by polarity: polar amino acids include arginine (basicpolar), asparagine, aspartic acid (acidic polar), glutamic acid (acidicpolar), glutamine, histidine (basic polar), lysine (basic polar),serine, threonine, and tyrosine; non-polar amino acids include alanine,cysteine, glycine, isoleucine, leucine, methionine, phenylalanine,proline, tryptophan, and valine. Thus, one or more amino acid residueswithin a CDR region can be replaced with other amino acid residues fromthe same group and the altered antibody can be tested for retainedfunction (i.e., the functions set forth in (c) through (l) above) usingthe functional assays described herein. In certain embodiments, no morethan one, no more than two, no more than three, no more than four, nomore than five residues within a specified sequence or a CDR region arealtered. Exemplary conservative amino acid substitutions are shown inTable 18.

TABLE 18 Exemplary conservative Original Residue amino acidSubstitutions Ala (A) Val; Leu; Ile Arg (R) Lys; Gln; Asn Asn (N) Gln;His; Asp, Lys; Arg Asp (D) Glu; Asn Cys (C) Ser; Ala Gln (Q) Asn; GluGlu (E) Asp; Gln Gly (G) Ala His (H) Asn; Gln; Lys; Arg Ile (I) Leu;Val; Met; Ala; Phe Leu (L) Ile; Val; Met; Ala; Phe Lys (K) Arg; Gln; AsnMet (M) Leu; Phe; Ile Phe (F) Trp; Leu; Val; Ile; Ala; Tyr Pro (P) AlaSer (S) Thr Thr (T) Val; Ser Trp (W) Tyr; Phe Tyr (Y) Trp; Phe; Thr; SerVal (V) Ile; Leu; Met; Phe; Ala

In certain non-limiting embodiments, an extracellular antigen-bindingdomain of the CAR can comprise a linker connecting the heavy chainvariable region and light chain variable region of the extracellularantigen-binding domain. As used herein, the term “linker” refers to afunctional group (e.g., chemical or polypeptide) that covalentlyattaches two or more polypeptides or nucleic acids so that they areconnected to one another. As used herein, a “peptide linker” refers toone or more amino acids used to couple two proteins together (e.g., tocouple V_(H) and V_(L) domains). Non-limiting examples of peptidelinkers are disclosed in Shen et al., Anal. Chem. 80(6):1910-1917 (2008)and WO 2014/087010.

In one non-limiting example, the linker comprises amino acids having thesequence set forth in SEQ ID NO:69. In certain embodiments, thenucleotide sequence encoding the amino acid sequence of SEQ ID NO:69 isset forth in SEQ ID NO:70. In one non-limiting example, the linker is aG45 linker that comprises amino acids having the sequence set forth inSEQ ID NO:210. In certain embodiments, the nucleotide sequence encodingthe amino acid sequence of SEQ ID NO:98 is set forth in SEQ ID NO:211.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:212 as provided below.

[SEQ ID NO: 212] GGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:213 as provided below.

[SEQ ID NO: 213] SGGSGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:214 as provided below.

[SEQ ID NO: 214] GGGGSGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:215 as provided below.

[SEQ ID NO: 215] GGGGSGGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:216 as provided below.

[SEQ ID NO: 216] GGGGSGGGGSGGGGGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:217 as provided below.

[SEQ ID NO: 217] GGGGSGGGGSGGGGSGGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:218 as provided below.

[SEQ ID NO: 218] GGGGSGGGGSGGGGSGGGGSGGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:219 as provided below.

[SEQ ID NO: 219] GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:220 as provided below.

[SEQ ID NO: 220] GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:220 as provided below.

[SEQ ID NO: 220] EPKSCDKTHTCPPCP.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:221 as provided below.

[SEQ ID NO: 222] GGGGSGGGSEPKSCDKTHTCPPCP.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:223 as provided below.

[SEQ ID NO: 223] ELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPEP KSCDTPPPCPRCP

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:224 as provided below.

[SEQ ID NO: 224] GSGSGS.

In certain embodiments, the linker comprises amino acids having thesequence set forth in SEQ ID NO:225 as provided below.

[SEQ ID NO: 225] AAA.

In addition, the extracellular antigen-binding domain can comprise aleader or a signal peptide that directs the nascent protein into theendoplasmic reticulum. Signal peptide or leader can be essential if theCAR is to be glycosylated and anchored in the cell membrane. The signalsequence or leader can be a peptide sequence (about 5, about 10, about15, about 20, about 25, or about 30 amino acids long) present at theN-terminus of newly synthesized proteins that directs their entry to thesecretory pathway. In non-limiting examples, the signal peptide iscovalently joined to the 5′ terminus of the extracellularantigen-binding domain. In certain embodiments, the signal peptidecomprises a CD8 polypeptide comprising amino acids having the sequenceset forth in SEQ ID NO:191 as provided below.

[SEQ ID NO: 191] MALPVTALLLPLALLLHAAR

The nucleotide sequence encoding the amino acid sequence of SEQ IDNO:191 is set forth in SEQ ID NO:192, which is provided below:

[SEQ ID NO: 192] ATGGCTCTCCCAGTGACTGCCCTACTGCTTCCCCTAGCGCTTCTCCTGCATGCAGCTCGT

In another embodiment, the signal peptide comprises amino acids havingthe sequence set forth in SEQ ID NO:205 as provided below.

[SEQ ID NO: 205] METDTLLLWVLLLWVPGSTGThe nucleotide sequence encoding the amino acid sequence of SEQ IDNO:205 is set forth in SEQ ID NO:206, which is provided below:

[SEQ ID NO: 206] ATGGAAACCGACACCCTGCTGCTGTGGGTGCTGCTGCTGTGGGTG CCAGGATCCACAGGA

In certain embodiments, the human scFv comprises a heavy chain variableregion, a light chain variable region, a linker peptide between theheavy chain variable region and the light chain variable region, and anHis-tag and an HA-tag. In certain embodiments, the amino acid sequenceof the His-tag and HA-tag comprises the amino acid sequence of SEQ IDNO:275, which is provided below:

[SEQ ID NO: 275] TSGQAGQHHHHHHGAYPYDVPDYAS

The nucleotide sequence encoding SEQ ID NO: 275 is SEQ ID NO: 276, whichis provided below:

[SEQ ID NO: 276] ACTAGTGGCCAGGCCGGCCAGCACCATCACCATCACCATGGCGCATACCCGTACGACGTTCCGGACTACGCTTCT

In certain embodiments, the extracellular antigen-binding domain (e.g.,a human scFv) binds to a human BCMA polypeptide comprising the aminoacid sequence set forth in SEQ ID NO: 71. In certain embodiments, theextracellular antigen-binding domain (e.g., a human scFv) binds to oneor more portion of the amino acid sequence set forth in SEQ ID NO: 71.In certain embodiments, the extracellular antigen-binding domain (e.g.,a human scFv) binds to an epitope region comprising amino acids 14-22 ofSEQ ID NO: 71. In certain embodiments, the extracellular antigen-bindingdomain (e.g., a human scFv) binds to one, two, three, four, five, six,or seven epitope region selected from the group consisting of aminoacids 8-22, 9-23, 10-24, 11-25, 12-26, 13-27, 14-28 and 8-28 of SEQ IDNO: 71. In certain embodiments, the extracellular antigen-binding domain(e.g., a human scFv) that binds to an epitope region comprising aminoacids 14-22 of SEQ ID NO: 71 comprises a heavy chain variable regioncomprising amino acids having the sequence set forth in SEQ ID NO:9 anda light chain variable region comprising amino acids having the sequenceset forth in SEQ ID NO:10, optionally with (iii) a linker sequence, forexample a linker peptide, between the heavy chain variable region andthe light chain variable region. In certain embodiments, the linkercomprises amino acids having the sequence set forth in SEQ ID NO:69. Incertain embodiments, the extracellular antigen-binding domain that bindsto an epitope region comprising amino acids 14-22 of SEQ ID NO: 71 is ahuman scFv with V_(H) and V_(L) regions or CDRs selected from Table 6.In certain embodiments, the extracellular antigen-binding domain thatbinds to amino acids 14-22 of SEQ ID NO: 71 comprises is a human scFv-Fcfusion protein or full length human IgG with V_(H) and V_(L) regions orCDRs selected from Table 6. In certain embodiments, the extracellularantigen-binding domain (e.g., a human scFv) that binds to an epitoperegion comprising amino acids 14-22 of SEQ ID NO: 71 comprises a V_(H)comprising an amino acid sequence that is at least 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% homologous to the amino acid sequence set forth in SEQ IDNO:21. In certain embodiments, the extracellular antigen-binding domain(e.g., a human scFv) that binds to an epitope region comprising aminoacids 14-22 of SEQ ID NO: 71 comprises a V_(H) comprising amino acidshaving the sequence set forth in SEQ ID NO:21. In certain embodiments,the extracellular antigen-binding domain (e.g., a human scFv) that bindsto an epitope region comprising amino acids 14-22 of SEQ ID NO: 71comprises a V_(L) comprising an amino acid sequence that is at least80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% or 99% homologous to the amino acid sequence setforth in SEQ ID NO:22. In certain embodiments, the extracellularantigen-binding domain (e.g., a human scFv) that binds to an epitoperegion comprising amino acids 14-22 of SEQ ID NO: 71 comprises a V_(L)comprising amino acids having the sequence set forth in SEQ ID NO:22. Incertain embodiments, the extracellular antigen-binding domain (e.g., ahuman scFv) that binds to an epitope region comprising amino acids 14-22of SEQ ID NO: 71 comprises a V_(H) comprising amino acids having thesequence set forth in SEQ ID NO:21 and a V_(L) comprising amino acidshaving the sequence set forth in SEQ ID NO:22. In certain embodiments,the extracellular antigen-binding domain (e.g., a human scFv) that bindsto an epitope region comprising amino acids 14-22 of SEQ ID NO: 71comprises a V_(H) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO:119 or conservative modifications thereof, a V_(H)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:120 or conservative modifications thereof, and a V_(H) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:121 orconservative modifications thereof. In certain embodiments, theextracellular antigen-binding domain (e.g., a human scFv) that binds toan epitope region comprising amino acids 14-22 of SEQ ID NO: 71comprises a V_(L) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO:122 or conservative modifications thereof, a V_(L)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:123 or conservative modifications thereof, and a V_(L) CDR3comprising amino acids having the sequence set forth in SEQ ID NO:124 orconservative modifications thereof. In certain embodiments, theextracellular antigen-binding domain (e.g., a human scFv) that binds toan epitope region comprising amino acids 14-22 of SEQ ID NO: 71comprises a V_(H) CDR1 comprising amino acids having the sequence setforth in SEQ ID NO:119 or conservative modifications thereof, a V_(H)CDR2 comprising amino acids having the sequence set forth in SEQ IDNO:120 or conservative modifications thereof, a V_(H) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO:121 orconservative modifications thereof, a V_(L) CDR1 comprising amino acidshaving the sequence set forth in SEQ ID NO:122 or conservativemodifications thereof, a V_(L) CDR2 comprising amino acids having thesequence set forth in SEQ ID NO:123 or conservative modificationsthereof, and a V_(L) CDR3 comprising amino acids having the sequence setforth in SEQ ID NO:124 or conservative modifications thereof. In certainembodiments, the extracellular antigen-binding domain (e.g., a humanscFv) that binds to an epitope region comprising amino acids 14-22 ofSEQ ID NO: 71 comprises a V_(H) CDR1 comprising amino acids having thesequence set forth in SEQ ID NO:119, a V_(H) CDR2 comprising amino acidshaving the sequence set forth in SEQ ID NO:120, a V_(H) CDR3 comprisingamino acids having the sequence set forth in SEQ ID NO:121, a V_(L) CDR1comprising amino acids having the sequence set forth in SEQ ID NO:122, aV_(L) CDR2 comprising amino acids having the sequence set forth in SEQID NO:123, and a V_(L) CDR3 comprising amino acids having the sequenceset forth in SEQ ID NO:124. In certain embodiments, the extracellularantigen-binding domain is ET140-3 (or “ET140-153”) scFv.

Transmembrane Domain of a CAR

In certain non-limiting embodiments, the transmembrane domain of the CARcomprises a hydrophobic alpha helix that spans at least a portion of themembrane. Different transmembrane domains result in different receptorstability. After antigen recognition, receptors cluster and a signal istransmitted to the cell. In accordance with the presently disclosedsubject matter, the transmembrane domain of the CAR can comprise a CD8polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide,a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide,a BTLA polypeptide, a synthetic peptide (not based on a proteinassociated with the immune response), or a combination thereof.

In certain embodiments, the transmembrane domain of a presentlydisclosed CAR comprises a CD28 polypeptide. The CD28 polypeptide canhave an amino acid sequence that is at least about 85%, about 90%, about95%, about 96%, about 97%, about 98%, about 99% or 100% homologous tothe sequence having a NCBI Reference No: P10747 or NP 006130 (SEQ IDNo:193), or fragments thereof, and/or may optionally comprise up to oneor up to two or up to three conservative amino acid substitutions. Incertain embodiments, the CD28 polypeptide can have an amino acidsequence that is a consecutive portion of SEQ ID NO: 193 which is atleast 20, or at least 30, or at least 40, or at least 50, and up to 220amino acids in length. Alternatively or additionally, in non-limitingvarious embodiments, the CD28 polypeptide has an amino acid sequence ofamino acids 1 to 220, 1 to 50, 50 to 100, 100 to 150, 150 to 200, or 200to 220 of SEQ ID NO: 193. In certain embodiments, the CAR of thepresently disclosed comprises a transmembrane domain comprising a CD28polypeptide, and an intracellular domain comprising a co-stimulatorysignaling region that comprises a CD28 polypeptide. In certainembodiments, the CD28 polypeptide comprised in the transmembrane domainand the intracellular domain has an amino acid sequence of amino acids114 to 220 of SEQ ID NO: 193.

SEQ ID NO: 193 is provided below:

[SEQ ID NO: 193]   1 MLRLLLALNL FPSIQVTGNK ILVKQSPMLV AYDNAVNLSC     KYSYNLFSRE FRASLHKGLD 61 SAVEVCVVYG NYSQQLQVYS KTGFNCDGKL GNESVTFYLQ     NLYVNQTDIY FCKIEVMYPP121 PYLDNEKSNG TIIHVKGKHL CPSPLFPGPS KPFWVLVVVG     GVLACYSLLV TVAFIIFWVR181 SKRSRLLHSD YMNMTPRRPG PTRKHYQPYA PPRDFAAYRS 

In accordance with the presently disclosed subject matter, a “CD28nucleic acid molecule” refers to a polynucleotide encoding a CD28polypeptide. In certain embodiments, the CD28 nucleic acid moleculeencoding the CD28 polypeptide comprised in the transmembrane domain andthe intracellular domain (e.g., the co-stimulatory signaling region) ofthe presently disclosed CAR (amino acids 114 to 220 of SEQ ID NO: 193)comprises nucleic acids having the sequence set forth in SEQ ID NO: 194as provided below.

[SEQ ID NO: 194] ATTGAAGTTATGTATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACCTTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCC

In certain embodiments, the transmembrane domain of a presentlydisclosed CAR comprises a CD8 polypeptide. The CD8 polypeptide can havean amino acid sequence that is at least about 85%, about 90%, about 95%,about 96%, about 97%, about 98%, about 99% or 100% homologous to thesequence having a NCBI Reference No: AAH25715 (SEQ ID No:226), orfragments thereof, and/or may optionally comprise up to one or up to twoor up to three conservative amino acid substitutions. In non-limitingembodiments, the CD8 polypeptide can have an amino acid sequence that isa consecutive portion of SEQ ID NO: 226 which is at least 20, or atleast 30, or at least 40, or at least 50, or at least 70, or at least100, or at least 150, or at least 200 and up to 235 amino acids inlength. Alternatively or additionally, in non-limiting variousembodiments, the CD28 polypeptide has an amino acid sequence of aminoacids 1 to 235, 1 to 50, 50 to 100, 100 to 150, 150 to 200, 130 to 210,or 200 to 235 of SEQ ID NO: 226. In certain embodiments, the CD8polypeptide comprised in the transmembrane domain has an amino acidsequence of amino acids 137 to 207 of SEQ ID NO: 226.

SEQ ID NO: 226 is provided below:

[SEQ ID NO: 226]   1 MALPVTALLL PLALLLHAAR PSQFRVSPLD RTWNLGETVE      LKCQVLLSNP TSGCSWLFQP 61 RGAAASPTFL LYLSQNKPKA AEGLDTQRFS GKRLGDTFVL    TLSDFRRENE GCYFCSALSN121 SIMYFSHFVP VFLPAKPTTT PAPRPPTPAP TIASQPLSLR     PEACRPAAGG AVHTRGLDFA181 CDIYIWAPLA GTCGVLLLSL VITLYCNHRN RRRVCKCPRP       VVKSGDKPSL SARYV.

In accordance with the presently disclosed subject matter, a “CD8nucleic acid molecule” refers to a polynucleotide encoding a CD8polypeptide. In certain embodiments, the CD8 nucleic acid moleculeencoding the CD8 polypeptide comprised in the transmembrane domain ofthe presently disclosed CAR (amino acids 137 to 207 of SEQ ID NO: 226)comprises nucleic acids having the sequence set forth in SEQ ID NO: 227as provided below.

[SEQ ID NO: 227] CCCACCACGACGCCAGCGCCGCGACCACCAACCCCGGCGCCCACGATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCGCCTGTGATATCTACATCTGGGCGCCCCTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTGGTTATCACCCTTTACTGCAAC 

In certain non-limiting embodiments, a CAR can also comprise a spacerregion that links the extracellular antigen-binding domain to thetransmembrane domain. The spacer region can be flexible enough to allowthe antigen binding domain to orient in different directions tofacilitate antigen recognition. The spacer region can be the hingeregion from IgG1, or the CH₂CH₃ region of immunoglobulin and portions ofCD3.

Intracellular Domain of a CAR

In certain non-limiting embodiments, an intracellular domain of the CARcan comprise a CD3ζ polypeptide, which can activate or stimulate a cell(e.g., a cell of the lymphoid lineage, e.g., a T cell). CD3ζ comprisesthree ITAMs, and transmits an activation signal to the cell (e.g., acell of the lymphoid lineage, e.g., a T cell) after antigen is bound.The CD3ζ polypeptide can have an amino acid sequence that is at leastabout 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about99% or about 100% homologous to the sequence set forth in SEQ ID NO:195, or fragments thereof, and/or may optionally comprise up to one orup to two or up to three conservative amino acid substitutions. Innon-limiting embodiments, the CD3ζ polypeptide can have an amino acidsequence that is a consecutive portion of SEQ ID NO: 195 which is atleast 20, or at least 30, or at least 40, or at least 50, and up to 163amino acids in length. Alternatively or additionally, in non-limitingvarious embodiments, the CD3ζ polypeptide has an amino acid sequence ofamino acids 1 to 163, 1 to 50, 50 to 100, 100 to 150, or 150 to 163 ofSEQ ID NO: 195. In certain embodiments, the CD3ζ polypeptide comprisedin the intracellular domain of a presently disclosed CAR has an aminoacid sequence of amino acids 52 to 163 of SEQ ID NO: 195.

SEQ ID NO: 195 is provided below:

[SEQ ID NO: 195]   1 MKWKALFTAA ILQAQLPITE AQSFGLLDPK LCYLLDGILF     IYGVILTALF LRVKFSRSAD 61 APAYQQGQNQ LYNELNLGRR EEYDVLDKRR GRDPEMGGKP     RRKNPQEGLY NELQKDKMAE121 AYSEIGMKGE RRRGKGHDGL YQGLSTATKD TYDALHMQAL       PPR

In accordance with the presently disclosed subject matter, a “CD3ζnucleic acid molecule” refers to a polynucleotide encoding a CD3ζpolypeptide. In certain embodiments, the CD3ζ nucleic acid moleculeencoding the CD3ζ polypeptide comprised in the intracellular domain of apresently disclosed CARs (amino acid sequence of amino acids 52 to 163of SEQ ID NO: 195) comprises nucleic acids having the sequence set forthin SEQ ID NO:196 as provided below.

[SEQ ID NO: 196] AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGG CCCTGCCCCCTCGCTAA

In certain non-limiting embodiments, an intracellular domain of the CARfurther comprises at least one signaling region. The at least onesignaling region can include a CD28 polypeptide, a 4-1BB polypeptide, anOX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, a PD-1polypeptide, a CTLA-4 polypeptide, a LAG-3 polypeptide, a 2B4polypeptide, a BTLA polypeptide, a synthetic peptide (not based on aprotein associated with the immune response), or a combination thereof.

In certain embodiments, the signaling region is a co-stimulatorysignaling region. In certain embodiments, the co-stimulatory regioncomprises at least one co-stimulatory molecule, which can provideoptimal lymphocyte activation. As used herein, “co-stimulatorymolecules” refer to cell surface molecules other than antigen receptorsor their ligands that are required for an efficient response oflymphocytes to antigen. The at least one co-stimulatory signaling regioncan include a CD28 polypeptide, a 4-1BB polypeptide, an OX40polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combinationthereof. The co-stimulatory molecule can bind to a co-stimulatoryligand, which is a protein expressed on cell surface that upon bindingto its receptor produces a co-stimulatory response, i.e., anintracellular response that effects the stimulation provided when anantigen binds to its CAR molecule. Co-stimulatory ligands, include, butare not limited to CD80, CD86, CD70, OX40L, 4-1BBL, CD48, TNFRSF14, andPD-L1. As one example, a 4-1BB ligand (i.e., 4-1BBL) may bind to 4-1BB(also known as “CD137”) for providing an intracellular signal that incombination with a CAR signal induces an effector cell function of theCAR⁺ T cell. CARs comprising an intracellular domain that comprises aco-stimulatory signaling region comprising 4-1BB, ICOS or DAP-10 aredisclosed in U.S. Pat. No. 7,446,190 (e.g., the nucleotide sequenceencoding 4-1BB is set forth in SEQ ID NO:15, the nucleotide sequenceencoding ICOS is set forth in SEQ ID NO:16, and the nucleotide sequenceencoding DAP-10 is set forth in SEQ ID NO:17 in U.S. Pat. No.7,446,190), which is herein incorporated by reference in its entirety.In certain embodiments, the intracellular domain of the CAR comprises aco-stimulatory signaling region that comprises a CD28 polypeptide. Incertain embodiments, the intracellular domain of the CAR comprises aco-stimulatory signaling region that comprises two co-stimulatorymolecules:CD28 and 4-1BB or CD28 and OX40.

4-1BB can act as a tumor necrosis factor (TNF) ligand and havestimulatory activity. The 4-1BB polypeptide can have an amino acidsequence that is at least about 85%, about 90%, about 95%, about 96%,about 97%, about 98%, about 99% or 100% homologous to the sequencehaving a NCBI Reference No: P41273 or NP 001552 (SEQ ID NO:197) orfragments thereof, and/or may optionally comprise up to one or up to twoor up to three conservative amino acid substitutions. In certainembodiments, the 4-1BB polypeptide comprised in the intracellular domainof a presently disclosed CAR has an amino acid sequence of amino acids214-255 of SEQ ID NO: 197.

SEQ ID NO: 197 is provided below:

[SEQ ID NO: 197]   1 MGNSCYNIVA TLLLVLNFER TRSLQDPCSN CPAGTFCDNN     RNQICSPCPP NSFSSAGGQR 61 TCDICRQCKG VFRTRKECSS TSNAECDCTP GFHCLGAGCS     MCEQDCKQGQ ELTKKGCKDC121 CFGTFNDQKR GICRPWTNCS LDGKSVLVNG TKERDVVCGP     SPADLSPGAS SVTPPAPARE181 PGHSPQIISF FLALTSTALL FLLFFLTLRF SVVKRGRKKL     LYIFKQPFMR PVQTTQEEDG 241 CSCRFPEEEE GGCEL 

In accordance with the presently disclosed subject matter, a “4-1BBnucleic acid molecule” refers to a polynucleotide encoding a 4-1BBpolypeptide. In certain embodiments, the 4-1BB nucleic acid moleculeencoding the 4-1BB polypeptide comprised in the intracellular domain ofa presently disclosed CARs (amino acids 214-255 of SEQ ID NO: 197)comprises nucleic acids having the sequence set forth in SEQ ID NO: 228as provided below.

[SEQ ID NO: 228] aaacggggcagaaagaagctcctgtatatattcaaacaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactg

An OX40 polypeptide can have an amino acid sequence that is at leastabout 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about99% or 100% homologous to the sequence having a NCBI Reference No:P43489 or NP 003318 (SEQ ID NO: 198), or fragments thereof, and/or mayoptionally comprise up to one or up to two or up to three conservativeamino acid substitutions.

SEQ ID NO: 198 is provided below:

[SEQ ID NO: 198]   1 MCVGARRLGR GPCAALLLLG LGLSTVTGLH CVGDTYPSND     RCCHECRPGN GMVSRCSRSQ 61 NTVCRPCGPG FYNDVVSSKP CKPCTWCNLR SGSERKQLCT     ATQDTVCRCR AGTQPLDSYK121 PGVDCAPCPP GHFSPGDNQA CKPWTNCTLA GKHTLQPASN     SSDAICEDRD PPATQPQETQ181 GPPARPITVQ PTEAWPRTSQ GPSTRPVEVP GGRAVAAILG     LGLVLGLLGP LAILLALYLL 241 RRDQRLPPDA HKPPGGGSFR TPIQEEQADA HSTLAKI 

In accordance with the presently disclosed subject matter, an “OX40nucleic acid molecule” refers to a polynucleotide encoding an OX40polypeptide.

An ICOS polypeptide can have an amino acid sequence that is at leastabout 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about99% or 100% homologous to the sequence having a NCBI Reference No:NP_036224 (SEQ ID NO: 199) or fragments thereof, and/or may optionallycomprise up to one or up to two or up to three conservative amino acidsubstitutions.

SEQ ID NO: 199 is provided below:

[SEQ ID NO: 199]   1 MKSGLWYFFL FCLRIKVLTG EINGSANYEM FIFHNGGVQI     LCKYPDIVQQ FKMQLLKGGQ 61 ILCDLTKTKG SGNTVSIKSL KFCHSQLSNN SVSFFLYNLD     HSHANYYFCN LSIFDPPPFK121 VTLIGGYLHI YESQLCCQLK FWLPIGCAAF VVVCILGCIL     ICWLTKKKYS SSVHDPNGEY 181 MFMRAVNTAK KSRLTDVTL

In accordance with the presently disclosed subject matter, an “ICOSnucleic acid molecule” refers to a polynucleotide encoding an ICOSpolypeptide.

CTLA-4 is an inhibitory receptor expressed by activated T cells, whichwhen engaged by its corresponding ligands (CD80 and CD86; B7-1 and B7-2,respectively), mediates activated T cell inhibition or anergy. In bothpreclinical and clinical studies, CTLA-4 blockade by systemic antibodyinfusion, enhanced the endogenous anti-tumor response albeit, in theclinical setting, with significant unforeseen toxicities.

CTLA-4 contains an extracellular V domain, a transmembrane domain, and acytoplasmic tail. Alternate splice variants, encoding differentisoforms, have been characterized. The membrane-bound isoform functionsas a homodimer interconnected by a disulfide bond, while the solubleisoform functions as a monomer. The intracellular domain is similar tothat of CD28, in that it has no intrinsic catalytic activity andcontains one YVKM motif able to bind PI3K, PP2A and SHP-2 and oneproline-rich motif able to bind SH3 containing proteins. One role ofCTLA-4 in inhibiting T cell responses seem to be directly via SHP-2 andPP2A dephosphorylation of TCR-proximal signaling proteins such as CD3and LAT. CTLA-4 can also affect signaling indirectly via competing withCD28 for CD80/86 binding. CTLA-4 has also been shown to bind and/orinteract with PI3K, CD80, AP2M1, and PPP2R5A.

In accordance with the presently disclosed subject matter, a CTLA-4polypeptide can have an amino acid sequence that is at least about 85%,about 90%, about 95%, about 96%, about 97%, about 98%, about 99% orabout 100% homologous to UniProtKB/Swiss-Prot Ref. No.: P16410.3 (SEQ IDNO: 200) (homology herein may be determined using standard software suchas BLAST or FASTA) or fragments thereof, and/or may optionally compriseup to one or up to two or up to three conservative amino acidsubstitutions.

SEQ ID NO: 200 is provided below:

[SEQ ID NO: 200]   1 MACLGFQRHK AQLNLATRTW PCTLLFFLLF IPVFCKAMHV     AQPAVVLASS RGIASFVCEY 61 ASPGKATEVR VTVLRQADSQ VTEVCAATYM MGNELTFLDD     SICTGTSSGN QVNLTIQGLR121 AMDTGLYICK VELMYPPPYY LGIGNGTQIY VIDPEPCPDS     DFLLWILAAV SSGLFFYSFL181 LTAVSLSKML KKRSPLTTGV YVKMPPTEPE CEKQFQPYFI       PIN

In accordance with the presently disclosed subject matter, a “CTLA-4nucleic acid molecule” refers to a polynucleotide encoding a CTLA-4polypeptide.

PD-1 is a negative immune regulator of activated T cells upon engagementwith its corresponding ligands PD-L1 and PD-L2 expressed on endogenousmacrophages and dendritic cells. PD-1 is a type I membrane protein of268 amino acids. PD-1 has two ligands, PD-L1 and PD-L2, which aremembers of the B7 family. The protein's structure comprises anextracellular IgV domain followed by a transmembrane region and anintracellular tail. The intracellular tail contains two phosphorylationsites located in an immunoreceptor tyrosine-based inhibitory motif andan immunoreceptor tyrosine-based switch motif, that PD-1 negativelyregulates TCR signals. SHP-I and SHP-2 phosphatases bind to thecytoplasmic tail of PD-1 upon ligand binding. Upregulation of PD-L1 isone mechanism tumor cells may evade the host immune system. Inpre-clinical and clinical trials, PD-1 blockade by antagonisticantibodies induced anti-tumor responses mediated through the hostendogenous immune system.

In accordance with the presently disclosed subject matter, a PD-1polypeptide can have an amino acid sequence that is at least about 85%,about 90%, about 95%, about 96%, about 97%, about 98%, about 99% orabout 100% homologous to NCBI Reference No: NP_005009.2 (SEQ ID NO: 201)or fragments thereof, and/or may optionally comprise up to one or up totwo or up to three conservative amino acid substitutions.

SEQ ID NO: 201 is provided below:

[SEQ ID NO: 201]   1 MQIPQAPWPV VWAVLQLGWR PGWFLDSPDR PWNPPTFSPA     LLVVTEGDNA TFTCSFSNTS 61 ESFVLNWYRM SPSNQTDKLA AFPEDRSQPG QDCRFRVTQL     PNGRDFHMSV VRARRNDSGT121 YLCGAISLAP KAQIKESLRA ELRVTERRAE VPTAHPSPSP     RPAGQFQTLV VGVVGGLLGS181 LVLLVWVLAV ICSRAARGTI GARRTGQPLK EDPSAVPVFS     VDYGELDFQW REKTPEPPVP241 CVPEQTEYAT IVFPSGMGTS SPARRGSADG PRSAQPLRPE      DGHCSWPL 

In accordance with the presently disclosed subject matter, a “PD-1nucleic acid molecule” refers to a polynucleotide encoding a PD-1polypeptide.

Lymphocyte-activation protein 3 (LAG-3) is a negative immune regulatorof immune cells. LAG-3 belongs to the immunoglobulin (1 g) superfamilyand contains 4 extracellular Ig-like domains. The LAG3 gene contains 8exons. The sequence data, exon/intron organization, and chromosomallocalization all indicate a close relationship of LAG3 to CD4. LAG3 hasalso been designated CD223 (cluster of differentiation 223).

In accordance with the the presently disclosed subject matter, a LAG-3polypeptide can have an amino acid sequence that is at least about 85%,about 90%, about 95%, about 96%, about 97%, about 98%, about 99% orabout 100% homologous to UniProtKB/Swiss-Prot Ref. No.: P18627.5 (SEQ IDNO: 202) or fragments thereof, and/or may optionally comprise up to oneor up to two or up to three conservative amino acid substitutions.

SEQ ID NO: 202 is provided below:

[SEQ ID NO: 202]   1 MWEAQFLGLL FLQPLWVAPV KPLQPGAEVP VVWAQEGAPA     QLPCSPTIPL QDLSLLRRAG 61 VTWQHQPDSG PPAAAPGHPL APGPHPAAPS SWGPRPRRYT     VLSVGPGGLR SGRLPLQPRV121 QLDERGRQRG DFSLWLRPAR RADAGEYRAA VHLRDRALSC     RLRLRLGQAS MTASPPGSLR181 ASDWVILNCS FSRPDRPASV HWFRNRGQGR VPVRESPHHH     LAESFLFLPQ VSPMDSGPWG241 CILTYRDGFN VSIMYNLTVL GLEPPTPLTV YAGAGSRVGL     PCRLPAGVGT RSFLTAKWTP301 PGGGPDLLVT GDNGDFTLRL EDVSQAQAGT YTCHIHLQEQ     QLNATVTLAI ITVTPKSFGS361 PGSLGKLLCE VTPVSGQERF VWSSLDTPSQ RSFSGPWLEA     QEAQLLSQPW QCQLYQGERL421 LGAAVYFTEL SSPGAQRSGR APGALPAGHL LLFLILGVLS     LLLLVTGAFG FHLWRRQWRP481 RRFSALEQGI HPPQAQSKIE ELEQEPEPEP EPEPEPEPEP      EPEQL 

In accordance with the presently disclosed subject matter, a “LAG-3nucleic acid molecule” refers to a polynucleotide encoding a LAG-3polypeptide.

Natural Killer Cell Receptor 2B4 (2B4) mediates non-MHC restricted cellkilling on NK cells and subsets of T cells. To date, the function of 2B4is still under investigation, with the 2B4-S isoform believed to be anactivating receptor, and the 2B4-L isoform believed to be a negativeimmune regulator of immune cells. 2B4 becomes engaged upon binding itshigh-affinity ligand, CD48. 2B4 contains a tyrosine-based switch motif,a molecular switch that allows the protein to associate with variousphosphatases. 2B4 has also been designated CD244 (cluster ofdifferentiation 244).

In accordance with the presently disclosed subject matter, a 2B4polypeptide can have an amino acid sequence that is at least about 85%,about 90%, about 95%, about 96%, about 97%, about 98%, about 99% orabout 100% homologous to UniProtKB/Swiss-Prot Ref. No.: Q9BZW8.2 (SEQ IDNO: 203) or fragments thereof, and/or may optionally comprise up to oneor up to two or up to three conservative amino acid substitutions.

SEQ ID NO: 203 is provided below:

[SEQ ID NO: 203]   1 MLGQVVTLIL LLLLKVYQGK GCQGSADHVV SISGVPLQLQ     PNSIQTKVDS IAWKKLLPSQ 61 NGFHHILKWE NGSLPSNTSN DRFSFIVKNL SLLIKAAQQQ     DSGLYCLEVT SISGKVQTAT121 FQVFVFESLL PDKVEKPRLQ GQGKILDRGR CQVALSCLVS     RDGNVSYAWY RGSKLIQTAG181 NLTYLDEEVD INGTHTYTCN VSNPVSWESH TLNLTQDCQN     AHQEFRFWPF LVIIVILSAL241 FLGTLACFCV WRRKRKEKQS ETSPKEFLTI YEDVKDLKTR     RNHEQEQTFP GGGSTIYSMI301 QSQSSAPTSQ EPAYTLYSLI QPSRKSGSRK RNHSPSFNST     IYEVIGKSQP KAQNPARLSR 361 KELENFDVYS 

In accordance with the presently disclosed subject matter, a “2B4nucleic acid molecule” refers to a polynucleotide encoding a 2B4polypeptide.

B- and T-lymphocyte attenuator (BTLA) expression is induced duringactivation of T cells, and BTLA remains expressed on Th1 cells but notTh2 cells. Like PD1 and CTLA4, BTLA interacts with a B7 homolog, B7H4.However, unlike PD-1 and CTLA-4, BTLA displays T-Cell inhibition viainteraction with tumor necrosis family receptors (TNF-R), not just theB7 family of cell surface receptors. BTLA is a ligand for tumor necrosisfactor (receptor) superfamily, member 14 (TNFRSF14), also known asherpes virus entry mediator (HVEM). BTLA-HVEM complexes negativelyregulate T-cell immune responses. BTLA activation has been shown toinhibit the function of human CD8⁺ cancer-specific T cells. BTLA hasalso been designated as CD272 (cluster of differentiation 272).

In accordance with the presently disclosed subject matter, a BTLApolypeptide can have an amino acid sequence that is at least about 85%,about 90%, about 95%, about 96%, about 97%, about 98%, about 99% orabout 100% homologous to UniProtKB/Swiss-Prot Ref. No.: Q7Z6A9.3 (SEQ IDNO: 204) or fragments thereof, and/or may optionally comprise up to oneor up to two or up to three conservative amino acid substitutions.

SEQ ID NO: 204 is provided below:

[SEQ ID NO: 204]   1 MKTLPAMLGT GKLFWVFFLI PYLDIWNIHG KESCDVQLYI     KRQSEHSILA GDPFELECPV 61 KYCANRPHVT WCKLNGTTCV KLEDRQTSWK EEKNISFFIL     HFEPVLPNDN GSYRCSANFQ121 SNLIESHSTT LYVTDVKSAS ERPSKDEMAS RPWLLYRLLP     LGGLPLLITT CFCLFCCLRR181 HQGKQNELSD TAGREINLVD AHLKSEQTEA STRQNSQVLL     SETGIYDNDP DLCFRMQEGS241 EVYSNPCLEE NKPGIVYASL NHSVIGPNSR LARNVKEAPT      EYASICVRS 

In accordance with the presently disclosed subject matter, a “BTLAnucleic acid molecule” refers to a polynucleotide encoding a BTLApolypeptide.

In certain embodiments, the CAR comprises an extracellularantigen-binding region that comprises a human scFv that specificallybinds to human BCMA, a transmembrane domain comprising a CD28polypeptide, and an intracellular domain comprising a CD3ζ polypeptideand a co-stimulatory signaling region that comprises a CD28 polypeptide,as shown in FIG. 1. As shown in FIG. 1, the CAR also comprises a signalpeptide or a leader covalently joined to the 5′ terminus of theextracellular antigen-binding domain. In certain embodiments, the signalpeptide comprises amino acids having the sequence set forth in SEQ IDNO:205.

In certain embodiments, the CAR comprises an extracellularantigen-binding region that comprises a human scFv that specificallybinds to human BCMA, a transmembrane domain comprising a CD8polypeptide, and an intracellular domain comprising a CD3ζ polypeptideand a co-stimulatory signaling region that comprises a 4-1BBpolypeptide, as shown in FIG. 7. As shown in FIG. 7, the CAR alsocomprises a signal peptide or a leader covalently joined to the 5′terminus of the extracellular antigen-binding domain. In certainembodiments, the signal peptide comprises amino acids having thesequence set forth in SEQ ID NO:205.

In certain embodiments, the CAR of the presently disclosed subjectmatter can further comprise an inducible promoter, for expressingnucleic acid sequences in human cells. Promoters for use in expressingCAR genes can be a constitutive promoter, such as ubiquitin C (UbiC)promoter.

The presently disclosed subject matter also provides isolated nucleicacid molecule encoding the BCMA-targeted CAR described herein or afunctional portion thereof. In certain embodiments, the isolated nucleicacid molecule encodes a presently disclosed BCMA-targeted CAR comprisinga human scFv that specifically binds to human BCMA, a transmembranedomain comprising a CD28 polypeptide, and an intracellular domaincomprising a CD3ξ polypeptide and a co-stimulatory signaling regioncomprising a CD28 polypeptide. In one specific non-limiting example, theisolated nucleic acid molecule comprises nucleic acids having thesequence set forth in SEQ ID NO:207 provided below:

[SEQ ID NO: 207] caatctgccctgactcagcctgcctccgtgtctgcgtctcctggacagtcgatcgccatctcctgcactggaaccagcagtgacgttggttggtatcaacagcacccaggcaaagcccccaaactcatgatttatgaggacagtaagcggccctcaggggrnctaatcgcttctctggctccaagtctggcaacacggcctccctgaccatctctgggctccaggctgaggacgaggctgattattactgcagctcaaatacaagaagcagcactttggtgttcggcggagggaccaagctgaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggccgaagtgcagctggtgcagtctggggctgagatgaagaagcctggggcctcactgaagctctcctgcaaggcttctggatacaccttcatcgactactatgtatactggatgcgacaggcccctggacaagggcttgagtccatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcagggcagggtcaccatgaccagggacacgtccatcagcacagcctacatggagctgagcaggctgagatctgacgacaccgccatgtattactgtgcgcgctcccagcgtgacggttacatggattactggggtcaaggtactctggtgaccgtctcctcagcggccgcaattgaagttatgtatcctcctccttacctagacaatgagaagagcaatggaaccattatccatgtgaaagggaaacacctttgtccaagtcccctatttcccggaccttctaagccgtttgggtgctggtggtggttggtggagtcctggcttgctatagcttgctagtaacagtggcctttattattttctgggtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccgggcccacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctccagagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgc 

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:208provided below:

[SEQ ID NO: 208] cagtctgtgctgacgcagccgccctcagtgtctggggccccagggcagagggtcaccatctcctgcactgggagcagctccaacatcggggcaggttttgatgtacactggtaccagcagcttccaggaacagcccccaaactcctcatctatggtaacagcaatcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcactgggctccaggctgaggatgaggctgattattactgccagtcctatgacagcagcctgagtggttatgtcttcggaactgggaccaaggtcaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggcccaggtccagctggtacagtctggggctgaggtgaagaagcctggggcctcagtgaaggtctcctgcaaggcttctggatacaccttcaccgactactatatgcactgggtgcgacaggcccctggacaacggcttgagtggatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcaggacaggatcaccgtgaccagggacacctccagcaacacaggctacatggagctgaccaggctgagatctgacgacacggccgtgtattactgtgcgcgctctccgtactctggtgttctggataaatggggtcaaggtactctggtgaccgtctcctcagcggccgcaattgaagttatgtatcctcctccttacctagacaatgagaagagcaatggaaccattatccatgtgaaagggaaacacctttgtccaagtcccctatttcccggaccttctaagcccttttgggtgctggtggtggttggtggagtcctggcttgctatagcttgctagtaacagtggcctttattattttctgggtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccgggcccacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctccagagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgcccgcggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccct

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:209provided below:

[SEQ ID NO: 209] tcctatgagctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatgtcttgttctggaaccagctccaacatcggaagtcactctgtaaactggtaccagcagctcccaggaacggcccccaaactcctcatctatactaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtggcctccagtctgaggatgaggctgattattactgtgcagcatgggatggcagcctgaatggtctggtattcggcggagggaccaagctgaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggccgaggtgcagctggtgcagtctggagcagaggtgaaaaagcccggggagtctctgaagatctcctgtaagggttctggatacagctttaccagctactggatcggctgggtgcgccagatgcccgggaaaggcctggagtggatggggatcatctatcctggtgactctgataccagatacagcccgtccttccaaggccacgtcaccatctcagctgacaagtccatcagcactgcctacctgcagtggagcagcctgaaggcctcggacaccgccatgtattactgtgcgcgctactctggttctttcgataactggggtcaaggtactctggtgaccgtctcctcagcggccgcaattgaagttatgtatcctcctccttacctagacaatgagaagagcaatggaaccattatccatgtgaaagggaaacacctttgtccaagtcccctatttcccggaccttctaagccctttgggtgctggtggtggttggtggagtcctggcttgctatagcttgctagtaacagtggcctttattattttctgggtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccgggcccacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctccagagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgc aggccctgccccctcgc 

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:229provided below:

[SEQ ID NO: 229] CCTTCTCTAGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCCCTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTCTACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCGACCGGTGCCGCCACCATGGAAACCGACACCCTGCTGCTGTGGGTGCTGCTGCTGTGGGTGCCAGGATCCACAGGACTGCCTGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGGACGCAGTTCCAACATCGGGAGTAATTCTGTTAACTGGTATCGACAACTCCCAGGAGCGGCCCCCAAACTCCTCATCTATAGTAATAATCAGCGGCCCCCAGGGGTCCCTGTGCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCAGTCTGAAGATGAGGCCACTTATTACTGTGCAACATGGGATGACAATCTGAATGTTCACTATGTCTTCGGAACTGGGACCAAGGTCACCGTCCTAGGTTCTAGAGGTGGTGGTGGTAGCGGCGGCGGCGGCTCTGGTGGTGGTGGATCCCTCGAGATGGCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAAGGATCATCCCTATCCTTGGTATAGCAAACTACGCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGCGCGGTGGTTACTACTCTCATGACATGTGGTCTGAAGATTGGGGTCAAGGTACTCTGGTGACCGTCTCCTCAGCggccgcacccaccacgacgccagcgccgcgaccaccaaccccggcgcccacgatcgcgtcgcagcccctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccctggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaacaaacggggcagaaagaagctcctgtatatattcaaacaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttgagcaggagcgcagagccccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaacagccactcgaggatccggattagtccaatttgttaaagacaggatatcagtggtccaggctctagttttgactcaacaatatcaccagctgaagcctatagagtacgagccatagataaaataaaagattttatttagtctccagaaaaaggggggaatgaaagaccccacctgtaggtttggcaagctagcttaagtaacgccattttgcaaggcatggaaaaatacataactgagaatagagaagttcagatcaaggtcaggaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggtccccagatgcggtccagccctcagcagtttctagagaaccatcagatgtttccagggtgccccaaggacctgaaatgaccctgtgccttatttgaactaaccaatcagttcgcttctcgcttctgttcgcgcgcttctgctccccgagctcaataaaagagcccacaacccctcactcggggcgccagtcctccgattgactgagtcgcccgggtacccgtgtatccaataaaccctcttgcagttgcatccgacttgtggtctcgctgttccttgggagggtctcctctgagtgattgactacccgtcagcgggggtctttcacacatgcagcatgtatcaaaattaatttggttttttttcttaagtatttacattaaatggccatagtacttaaagttacattggcttccttgaaataaacatggagtattcagaatgtgtcataaatatttctaattttaagatagtatctccattggctttctactttttcttttatttttttttgtcctctgtcttccatttgttgttgttgttgtttgtttgtttgtttgttggttggttggttaatttttttttaaagatcctacactatagttcaagctagactattagctactctgtaacccagggtgaccttgaagtcatgggtagcctgctgttttagccttcccacatctaagattacaggtatgagctatcatttttggtatattgattgattgattgattgatgtgtgtgtgtgtgattgtgtttgtgtgtgtgactgtgaaaatgtgtgtatgggtgtgtgtgaatgtgtgtatgtatgtgtgtgtgtgagtgtgtgtgtgtgtgtgtgcatgtgtgtgtgtgtgactgtgtctatgtgtatgactgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgttgtgaaaaaatattctatggtagtgagagccaacgctccggctcaggtgtcaggttggtttttgagacagagtctttcacttagcttggAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGATGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTTGCTCTTAGGAGTTTCCTAATACATCCCAAACTCAAATATATAAAGCATTTGACTTGTTCTATGCCCTAGGGGGCGGGGGGAAGCTAAGCCAGCTTTTTTTAACATTTAAAATGTTAATTCCATTTTAAATGCACAGATGTTTTTATTTCATAAGGGTTTCAATGTGCATGAATGCTGCAATATTCCTGTTACCAAAGCTAGTATAAATAAAAATAGATAAACGTGGAAATTACTTAGAGTTTCTGTCATTAACGTTTCCTTCCTCAGTTGACAACATAAATGCGCTGCTGAGCAAGCCAGTTTGCATCTGTCAGGATCAATTTCCCATTATGCCAGTCATATTAATTACTAGTCAATTAGTTGATTTTTATTTTTGACATATACATGTGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTGAGAATAGAAAAGTTCAGATCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGACCCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTCTGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAACTAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCCTGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCCGATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGACGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCCGCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATATGGGCCCGGGCTAGACTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCCAGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGGTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACT

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:230provided below:

[SEQ ID NO: 230] GGCCCTCTAGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCCCTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTCTACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCGACCGGTGCCGCCACCATGGAAACCGACACCCTGCTGCTGTGGGTGCTGCTGCTGTGGGTGCCAGGATCCACAGGAtcctatgagctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatgtcttgttctggaaccagctccaacatcggaagtcactctgtaaactggtaccagcagctcccaggaacggcccccaaactcctcatctatactaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtggcctccagtctgaggatgaggctgattattactgtgcagcatgggatggcagcctgaatggtctggtattcggcggagggaccaagctgaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggccgaggtgcagctggtgcagtctggagcagaggtgaaaaagcccggggagtctctgaagatctcctgtaagggttctggatacagctttaccagctactggatcggctgggtgcgccagatgcccgggaaaggcctggagtggatggggatcatctatcctggtgactctgataccagatacagcccgtccttccaaggccacgtcaccatctcagctgacaagtccatcagcactgcctacctgcagtggagcagcctgaaggcctcggacaccgccatgtattactgtgcgcgctactctggttctttcgataactggggtcaaggtactctggtgaccgtctcctcagcggccgcacccaccacgacgccagcgccgcgaccaccaaccccggcgcccacgatcgcgtcgcagcccctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccctggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaacaaacggggcagaaagaagctcctgtatatattcaaacaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcaggaggagcgcagagccccccgcgtaccaggagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaacagccactcgaggatccggattagtccaatttgttaaagacaggatatcagtggtccaggctctagttttgactcaacaatatcaccagctgaagcctatagagtacgagccatagataaaataaaagattttatttagtctccagaaaaaggggggaatgaaagaccccacctgtaggtttggcaagctagcttaagtaacgccattttgcaaggcatggaaaaatacataactgagaatagagaagttcagatcaaggtcaggaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggaacagctgaatatgggccaaacaggatatctgtggtaaggagttcctgccccggctcagggccaagaacagatggtccccagatgcggtccagccctcaggagtttctagagaaccatcagatgtttccagggtgccccaaggacctgaaatgaccctgtgccttatttgaactaaccaatcagttcgcttctcgcttctgttcgcgcgcttctgctccccgagctcaataaaagagcccacaacccctcactcggggcgccagtcctccgattgactgagtcgcccgggtacccgtgtatccaataaaccctcttgcagttgcatccgacttgtggtctcgctgttccttgggagggtctcctctgagtgattgactacccgtcagcgggggtctttcacacatgcagcatgtatcaaaattaatttggttttttttcttaagtatttacattaaatggccatagtacttaaagttacattggcttccttgaaataaacatggagtattcagaatgtgtcataaatatttctaattttaagatagtatctccattggctttctactttttcttttatttttttttgtcctctgtcttccatttgttgttgttgttgtttgtttgtttgtttgttggttggttggttaatttttttttaaagatcctacactatagttcaagctagactattagctactctgtaacccagggtgaccttgaagtcatgggtagcctgctgttttagccttcccacatctaagattacaggtatgagctatcatttttggtatattgattgattgattgattgatgtgtgtgtgtgtgattgtgtttgtgtgtgtgactgtgaaaatgtgtgtatgggtgtgtgtgaatgtgtgtatgtatgtgtgtgtgtgagtgtgtgtgtgtgtgtgtgcatgtgtgtgtgtgtgactgtgtctatgtgtatgactgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgttgtgaaaaaatattctatggtagtgagagccaacgctccggctcaggtgtcaggttggtttttgagacagagtctttcacttagcttggAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGATGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTTGCTCTTAGGAGTTTCCTAATACATCCCAAACTCAAATATATAAAGCATTTGACTTGTTCTATGCCCTAGGGGGCGGGGGGAAGCTAAGCCAGCTTTTTTTAACATTTAAAATGTTAATTCCATTTTAAATGCACAGATGTTTTTATTTCATAAGGGTTTCAATGTGCATGAATGCTGCAATATTCCTGTTACCAAAGCTAGTATAAATAAAAATAGTATAAACGTGGAAATTACTTAGAGTTTCTGTCATTAACGTTTCCTTCCTCAGTTGACAACATAAATGCGCTGCTGAGCAAGCCAGTTTGCATCTGTCAGGATCAATTTCCCATTATGCCAGTCATATTAATTACTAGTCAATTAGTTGATTTTTATTTTTGACATATACATGTGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTGAGAATAGAAAAGTTCAGATCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGACCCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTCTGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAACTAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCCTGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCCGATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGACGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCCGCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATATGGGCCCGGGCTAGACTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCCAGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGGTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCC TCCCTTTATCCAGCCCTCACT

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:231provided below:

[SEQ ID NO: 231] CCTTCTCTAGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCCCTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTCTACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCGACCGGTGCCGCCACCATGGAAACCGACACCCTGCTGCTGTGGGTGCTGCTGCTGTGGGTGCCAGGATCCACAGGACAGGCTGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATTACGTATTCTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGTAATAATCAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGACAGCCTGAGTGCCTCTTATGTTTTCGGAACTGGGACCAAGGTCACCGTCCTAGGTTCTAGAGGTGGTGGTGGTAGCGGCGGCGGCGGCTCTGGTGGTGGTGGATCCCTCGAGATGGCCCAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAAGGATCATCCCTATCCTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGCGCTCTGGTTACGGTTCTTACCGTTGGGAAGATTCTTGGGGTCAAGGTACTCTGGTGACCGTCTCCTCAGCggccgcacccaccacgacgccagcgccgcgaccaccaaccccggcgcccacgatcgcgtcgcagcccctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccctggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaacaaacggggcagaaagaagctcctgtatatattcaaacaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcaggaggagcgcagagccccccgcgtaccaggagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaacagccactcgaggatccggattagtccaatttgttaaagacaggatatcagtggtccaggctctagttttgactcaacaatatcaccagctgaagcctatagagtacgagccatagataaaataaaagattttatttagtctccagaaaaaggggggaatgaaagaccccacctgtaggtttggcaagctagcttaagtaacgccattttgcaaggcatggaaaaatacataactgagaatagagaagttcagatcaaggtcaggaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggtccccagatgcggtccagccctcagcagtttctagagaaccatcagatgtttccagggtgccccaaggacctgaaatgaccctgtgccttatttgaactaaccaatcagttcgcttctcgcttctgttcgcgcgcttctgctccccgagctcaataaaagagcccacaacccctcactcggggcgccagtcctccgattgactgagtcgcccgggtacccgtgtatccaataaaccctcttgcagttgcatccgacttgtggtctcgctgttccttgggagggtctcctctgagtgattgactacccgtcagcgggggtctttcacacatgcagcatgtatcaaaattaatttggttttttttcttaagtatttacattaaatggccatagtacttaaagttacattggcttccttgaaataaacatggagtattcagaatgtgtcataaatatttctaattttaagatagtatctccattggctttctactttttcttttatttttttttgtcctctgtcttccatttgttgttgttgttgtttgtttgtttgtttgttggttggttggttaatttttttttaaagatcctacactatagttcaagctagactattagctactctgtaacccagggtgaccttgaagtcatgggtagcctgctgttttagccttcccacatctaagattacaggtatgagctatcatttttggtatattgattgattgattgattgatgtgtgtgtgtgtgattgtgtttgtgtgtgtgactgtgaaaatgtgtgtatgggtgtgtgtgaatgtgtgtatgtatgtgtgtgtgtgagtgtgtgtgtgtgtgtgtgcatgtgtgtgtgtgtgactgtgtctatgtgtatgactgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgttgtgaaaaaatattctatggtagtgagagccaacgctccggctcaggtgtcaggttggtttttgagacagagtctttcacttagcttggAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGATGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTTGCTCTTAGGAGTTTCCTAATACATCCCAAACTCAAATATATAAAGCATTTGACTTGTTCTATGCCCTAGGGGGCGGGGGGAAGCTAAGCCAGCTTTTTTTAACATTTAAAATGTTAATTCCATTTTAAATGCACAGATGTTTTTATTTCATAAGGGTTTCAATGTGCATGAATGCTGCAATATTCCTGTTACCAAAGCTAGTATAAATAAAAATAGATAAACGTGGAAATTACTTAGAGTTTCTGTCATTAACGTTTCCTTCCTCAGTTGACAACATAAATGCGCTGCTGAGCAAGCCAGTTTGCATCTGTCAGGATCAATTTCCCATTATGCCAGTCATATTAATTACTAGTCAATTAGTTGATTTTTATTTTTGACATATACATGTGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTGAGAATAGAAAAGTTCAGATCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGACCCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTCTGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAACTAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCCTGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCCGATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGACGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCCGCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATATGGGCCCGGGCTAGACTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCCAGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGGTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACT 

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:232provided below:

[SEQ ID NO: 232] CCTTCTCTAGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCCCTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTCTACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCGACCGGTGCCGCCACCATGGAAACCGACACCCTGCTGCTGTGGGTGCTGCTGCTGTGGGTGCCAGGATCCACAGGAcagtctgtgctgacgcagccgccctcagtgtctggggccccagggcagagggtcaccatctcctgcactgggaggagctccaacatcggggcaggttttgatgtacactggtaccaggagcttccaggaacagcccccaaactcctcatctatggtaacagcaatcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcactgggctccaggctgaggatgaggctgattattactgccagtcctatgacagcagcctgagtggttatgtcttcggaactgggaccaaggtcaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggcccaggtccagctggtacagtctggggctgaggtgaagaagcctggggcctcagtgaaggtctcctgcaaggcttctggatacaccttcaccgactactatatgcactgggtgcgacaggcccctggacaacggcttgagtggatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcaggacaggatcaccgtgaccagggacacctccagcaacacaggctacatggagctgaccaggctgagatctgacgacacggccgtgtattactgtgcgcgctctccgtactctggtgttctggataaatggggtcaaggtactctggtgaccgtctcctcagcggccgcacccaccacgacgccagcgccgcgaccaccaaccccggcgcccacgatcgcgtcgcagcccctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccctggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaacaaacggggcagaaagaagctcctgtatatattcaaacaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcaggaggagcgcagagccccccgcgtaccaggagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaacagccactcgaggatccggattagtccaatttgttaaagacaggatatcagtggtccaggctctagttttgactcaacaatatcaccagctgaagcctatagagtacgagccatagataaaataaaagattttatttagtctccagaaaaaggggggaatgaaagaccccacctgtaggtttggcaagctagcttaagtaacgccattttgcaaggcatggaaaaatacataactgagaatagagaagttcagatcaaggtcaggaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggtccccagatgcggtccagccctcagcagtttctagagaaccatcagatgtttccagggtgccccaaggacctgaaatgaccctgtgccttatttgaactaaccaatcagttcgcttctcgcttctgttcgcgcgcttctgctccccgagctcaataaaagagcccacaacccctcactcggggcgccagtcctccgattgactgagtcgcccgggtacccgtgtatccaataaaccctcttgcagttgcatccgacttgtggtctcgctgttccttgggagggtctcctctgagtgattgactacccgtcagcgggggtctttcacacatgcagcatgtatcaaaattaatttggttttttttcttaagtatttacattaaatggccatagtacttaaagttacattggcttccttgaaataaacatggagtattcagaatgtgtcataaatatttctaattttaagatagtatctccattggctttctactttttcttttatttttttttgtcctctgtcttccatttgttgttgttgttgtttgtttgtttgtttgttggttggttggttaatttttttttaaagatcctacactatagttcaagctagactattagctactctgtaacccagggtgaccttgaagtcatgggtagcctgctgttttagccttcccacatctaagattacaggtatgagctatcatttttggtatattgattgattgattgattgatgtgtgtgtgtgtgattgtgtttgtgtgtgtgactgtgaaaatgtgtgtatgggtgtgtgtgaatgtgtgtatgtatgtgtgtgtgtgagtgtgtgtgtgtgtgtgtgcatgtgtgtgtgtgtgactgtgtctatgtgtatgactgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgttgtgaaaaaatattctatggtagtgagagccaacgctccggctcaggtgtcaggttggtttttgagacagagtctttcacttagcttggAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGATGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTTGCTCTTAGGAGTTTCCTAATACATCCCAAACTCAAATATATAAAGCATTTGACTTGTTCTATGCCCTAGGGGGCGGGGGGAAGCTAAGCCAGCTTTTTTTAACATTTAAAATGTTAATTCCATTTTAAATGCACAGATGTTTTTATTTCATAAGGGTTTCAATGTGCATGAATGCTGCAATATTCCTGTTACCAAAGCTAGTATAAATAAAAATAGATAAACGTGGAAATTACTTAGAGTTTCTGTCATTAACGTTTCCTTCCTCAGTTGACAACATAAATGCGCTGCTGAGCAAGCCAGTTTGCATCTGTCAGGATCAATTTCCCATTATGCCAGTCATATTAATTACTAGTCAATTAGTTGATTTTTATTTTTGACATATACATGTGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTGAGAATAGAAAAGTTCAGATCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGACCCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTCTGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAACTAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCCTGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCCGATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGACGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCCGCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATATGGGCCCGGGCTAGACTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCCAGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGGTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACT 

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:233provided below:

[SEQ ID NO: 233] CCTTCTCTAGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCCCTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTCTACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCGACCGGTGCCGCCACCATGGAAACCGACACCCTGCTGCTGTGGGTGCTGCTGCTGTGGGTGCCAGGATCCACAGGAcaatctgccctgactcagcctgcctccgtgtctgcgtctcctggacagtcgatcgccatctcctgcactggaaccagcagtgacgttggttggtatcaacagcacccaggcaaagcccccaaactcatgatttatgaggacagtaagcggccctcaggggtttctaatcgcttctctggctccaagtctggcaacacggcctccctgaccatctctgggctccaggctgaggacgaggctgattattactgcagctcaaatacaagaagcagcactttggtgttcggcggagggaccaagctgaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggccgaagtgcagctggtgcagtctggggctgagatgaagaagcctggggcctcactgaagctctcctgcaaggcttctggatacaccttcatcgactactatgtatactggatgcgacaggcccctggacaagggcttgagtccatgggatggatcaaccctaacagtggtggcacaaactatgcacagaagtttcagggcagggtcaccatgaccagggacacgtccatcagcacagcctacatggagctgagcaggctgagatctgacgacaccgccatgtattactgtgcgcgctcccagcgtgacggttacatggattactggggtcaaggtactctggtgaccgtctcctcagcggccgcacccaccacgacgccagcgccgcgaccaccaaccccggcgcccacgatcgcgtcgcagcccctgtccctgcgcccagaggcgtgccggccagcggcggggggcgcagtgcacacgagggggctggacttcgcctgtgatatctacatctgggcgcccctggccgggacttgtggggtccttctcctgtcactggttatcaccctttactgcaacaaacggggcagaaagaagctcctgtatatattcaaacaaccatttatgagaccagtacaaactactcaagaggaagatggctgtagctgccgatttccagaagaagaagaaggaggatgtgaactgagagtgaagttcaggaggagcgcagagccccccgcgtaccaggagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaacagccactcgaggatccggattagtccaatttgttaaagacaggatatcagtggtccaggctctagttttgactcaacaatatcaccagctgaagcctatagagtacgagccatagataaaataaaagattttatttagtctccagaaaaaggggggaatgaaagaccccacctgtaggtttggcaagctagcttaagtaacgccattttgcaaggcatggaaaaatacataactgagaatagagaagttcagatcaaggtcaggaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggaacagctgaatatgggccaaacaggatatctgtggtaagcagttcctgccccggctcagggccaagaacagatggtccccagatgcggtccagccctcagcagtttctagagaaccatcagatgtttccagggtgccccaaggacctgaaatgaccctgtgccttatttgaactaaccaatcagttcgcttctcgcttctgttcgcgcgcttctgctccccgagctcaataaaagagcccacaacccctcactcggggcgccagtcctccgattgactgagtcgcccgggtacccgtgtatccaataaaccctcttgcagttgcatccgacttgtggtctcgctgttccttgggagggtctcctctgagtgattgactacccgtcagcgggggtctttcacacatgcagcatgtatcaaaattaatttggttttttttcttaagtatttacattaaatggccatagtacttaaagttacattggcttccttgaaataaacatggagtattcagaatgtgtcataaatatttctaattttaagatagtatctccattggctttctactttttcttttatttttttttgtcctctgtcttccatttgttgttgttgttgtttgtttgtttgtttgttggttggttggttaatttttttttaaagatcctacactatagttcaagctagactattagctactctgtaacccagggtgaccttgaagtcatgggtagcctgctgttttagccttcccacatctaagattacaggtatgagctatcatttttggtatattgattgattgattgattgatgtgtgtgtgtgtgattgtgtttgtgtgtgtgactgtgaaaatgtgtgtatgggtgtgtgtgaatgtgtgtatgtatgtgtgtgtgtgagtgtgtgtgtgtgtgtgtgcatgtgtgtgtgtgtgactgtgtctatgtgtatgactgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgttgtgaaaaaatattctatggtagtgagagccaacgctccggctcaggtgtcaggttggtttttgagacagagtctttcacttagcttggAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGATGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTTGCTCTTAGGAGTTTCCTAATACATCCCAAACTCAAATATATAAAGCATTTGACTTGTTCTATGCCCTAGGGGGCGGGGGGAAGCTAAGCCAGCTTTTTTTAACATTTAAAATGTTAATTCCATTTTAAATGCACAGATGTTTTTATTTCATAAGGGTTTCAATGTGCATGAATGCTGCAATATTCCTGTTACCAAAGCTAGTATAAATAAAAATAGATAAACGTGGAAATTACTTAGAGTTTCTGTCATTAACGTTTCCTTCCTCAGTTGACAACATAAATGCGCTGCTGAGCAAGCCAGTTTGCATCTGTCAGGATCAATTTCCCATTATGCCAGTCATATTAATTACTAGTCAATTAGTTGATTTTTATTTTTGACATATACATGTGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTGAGAATAGAAAAGTTCAGATCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGACCCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTCTGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAACTAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCCTGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCCGATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGACGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCCGCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATATGGGCCCGGGCTAGACTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCCAGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGGTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCA  GCCCTCACT

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:234provided below:

[SEQ ID NO: 234] atggaaaccgacaccctgctgctgtgggtgctgctgctgtgggtgccaggatccacaggactgcctgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggacgcagttccaacatcgggagtaattctgttaactggtatcgacaactcccaggagcggcccccaaactcctcatctatagtaataatcagcggcccccaggggtccctgtgcgattctctggctccaagtctggcacctcagcctccctggccatcagtgggctccagtctgaagatgaggccacttattactgtgcaacatgggatgacaatctgaatgttcactatgtcttcggaactgggaccaaggtcaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggcccaggtgcagctggtgcagtctggggctgaggtgaagaagcctgggtcctcggtgaaggtctcctgcaaggcttctggaggcaccttcagcagctatgctatcagctgggtgcgacaggcccctggacaagggcttgagtggatgggaaggatcatccctatccttggtatagcaaactacgcacagaagttccagggcagagtcacgattaccgcggacaaatccacgagcacagcctacatggagctgagcagcctgagatctgaggacacggccgtgtattactgtgcgcgcggtggttactactctcatgacatgtggtctgaagattggggtcaaggtactctggtgaccgtctcctcagcggccgcaattgaagttatgtatcctcctccttacctagacaatgagaagagcaatggaaccattatccatgtgaaagggaaacacctttgtccaagtcccctatttcccggaccttctaagcccttttgggtgctggtggtggttggtggagtcctggcttgctatagcttgctagtaacagtggcctttattattttctgggtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccgggcccacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctccagagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaa

In one specific non-limiting example, the isolated nucleic acid moleculecomprises nucleic acids having the sequence set forth in SEQ ID NO:235provided below:

[SEQ ID NO: 235] atggaaaccgacaccctgctgctgtgggtgctgctgctgtgggtgccaggatccacaggacaggctgtgctgactcagccaccctcagcgtctgggacccccgggcagagggtcaccatctcttgttctggaagcagctccaacatcggaagtaattacgtattctggtaccagcagctcccaggaacggcccccaaactcctcatctatagtaataatcagcggccctcaggggtccctgaccgattctctggctccaagtctggcacctcagcctccctggccatcagtgggctccggtccgaggatgaggctgattattactgtgcagcatgggatgacagcctgagtgcctcttatgttttcggaactgggaccaaggtcaccgtcctaggttctagaggtggtggtggtagcggcggcggcggctctggtggtggtggatccctcgagatggcccaggtgcagctggtgcagtctggggctgaggtgaagaagcctgggtcctcggtgaaggtctcctgcaaggcttctggaggcaccttcagcagctatgctatcagctgggtgcgacaggcccctggacaagggcttgagtggatgggaaggatcatccctatccttggtacagcaaactacgcacagaagttccagggcagagtcacgattaccgcggacgaatccacgagcacagcctacatggagctgagcagcctgagatctgaggacacggccgtgtattactgtgcgcgctctggttacggttcttaccgttgggaagattcttggggtcaaggtactctggtgaccgtctcctcagcggccgcaattgaagttatgtatcctcctccttacctagacaatgagaagagcaatggaaccattatccatgtgaaagggaaacacctttgtccaagtcccctatttcccggaccttctaagcccttttgggtgctggtggtggttggtggagtcctggcttgctatagcttgctagtaacagtggcctttattattttctgggtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccgggcccacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctccagagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgctaa 

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:207 encodes a BCMA-targeted CAR (designated as BCMA-targeted 28zCAR54) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:53, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:54, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD28 polypeptide, and an intracellular domain comprising aCD3ξ polypeptide comprising amino acid sequence of amino acids 52 to 163of SEQ ID NO: 195, and a co-stimulatory signaling region comprising aCD28 polypeptide, wherein the CD28 region comprising the transmembrancedomain and the co-stimulatory signaling region comprises amino acids 114to 220 of SEQ ID NO:193.

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:208 encodes a BCMA-targeted CAR (designated as BCMA-targeted 28zCAR40) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:57, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:58, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD28 polypeptide, and an intracellular domain comprising aCD3ξ polypeptide comprising amino acid sequence of amino acids 52 to 163of SEQ ID NO: 195, and a co-stimulatory signaling region comprising aCD28 polypeptide, wherein the CD28 region comprising the transmembrancedomain and the co-stimulatory signaling region comprises amino acids 114to 220 of SEQ ID NO:193.

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:209 encodes a BCMA-targeted CAR (designated as BCMA-targeted 28zCAR24) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:65, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:66, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD28 polypeptide, and an intracellular domain comprising aCD3ξ polypeptide comprising amino acid sequence of amino acids 52 to 163of SEQ ID NO: 195, and a co-stimulatory signaling region comprising aCD28 polypeptide, wherein the CD28 region comprising the transmembrancedomain and the co-stimulatory signaling region comprises amino acids 114to 220 of SEQ ID NO:193.

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:234 encodes a BCMA-targeted CAR (designated as BCMA-targeted 28zCAR3) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:21, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:22, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD28 polypeptide, and an intracellular domain comprising aCD3ξ polypeptide comprising amino acid sequence of amino acids 52 to 163of SEQ ID NO: 195, and a co-stimulatory signaling region comprising aCD28 polypeptide, wherein the CD28 region comprising the transmembrancedomain and the co-stimulatory signaling region comprises amino acids 114to 220 of SEQ ID NO:193.

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:235 encodes a BCMA-targeted CAR (designated as BCMA-targeted 28zCAR37) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:61, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:62, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD28 polypeptide, and an intracellular domain comprising aCD3ξ polypeptide comprising amino acid sequence of amino acids 52 to 163of SEQ ID NO: 195, and a co-stimulatory signaling region comprising aCD28 polypeptide, wherein the CD28 region comprising the transmembrancedomain and the co-stimulatory signaling region comprises amino acids 114to 220 of SEQ ID NO:193.

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:229 encodes a BCMA-targeted CAR (designated as BCMA-targeted BBzCAR3) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:21, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:22, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD8 polypeptide having amino acids 137 to 207 of SEQ ID NO:226, and an intracellular domain comprising a CD3ξ polypeptidecomprising amino acid sequence of amino acids 52 to 163 of SEQ ID NO:195, and a co-stimulatory signaling region comprising a 4-1BBpolypeptide having the amino acids 214-255 of SEQ ID NO: 197. Nucleotidesequences 270-1031 of SEQ ID NO: 229 encodes the human scFv. Nucleotidesequences 1041-1253 of SEQ ID NO: 229 encodes the CD8 polypeptidecomprised in the transmembrane domain. Nucleotide sequences 1254-1379 ofSEQ ID NO: 229 encodes the 4-1BB polypeptide comprised in theintracellular domain. Nucleotide sequences 1380-1718 of SEQ ID NO: 229encodes the CD3zeta polypeptide comprised in the intracellular domain.Other portions of SEQ ID NO: 229 are shown in Table 19.

TABLE 19 nucleotide Sequence positions of SEQ number of Portions ID NO:229 nucleotides Kappa sp 210 . . . 269 60 LTR 1998 . . . 2467 470 M13fwd 3166 . . . 3182 17 AmpR 3657 . . . 3761 105 promoter AmpR 3762 . . .4622 861 ori 4793 . . . 5381 589 CAP binding 5669 . . . 5690 22 site lacpromoter 5705 . . . 5735 31 lac operator 5743 . . . 5759 17 M13 rev 5767. . . 5783 17 LTR 6192 . . . 6785 594 MMLV Psi 6848 . . . 7205 358 gag7270 . . . 15  417 (truncated)

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:230 encodes a BCMA-targeted CAR (designated as BCMA-targeted BBzCAR24) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:65, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:66, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD8 polypeptide having amino acids 137 to 207 of SEQ ID NO:226, and an intracellular domain comprising a CD3ξ polypeptidecomprising amino acid sequence of amino acids 52 to 163 of SEQ ID NO:195, and a co-stimulatory signaling region comprising a 4-1BBpolypeptide having the amino acids 214-255 of SEQ ID NO: 197. Nucleotidesequences 270-1015 of SEQ ID NO: 230 encodes the human scFv. Nucleotidesequences 1023-1235 of SEQ ID NO: 230 encodes the CD8 polypeptidecomprised in the transmembrane domain. Nucleotide sequences 1236-1361 ofSEQ ID NO: 230 encodes the 4-1BB polypeptide comprised in theintracellular domain. Nucleotide sequences 1362-1700 of SEQ ID NO: 230encodes the CD3zeta polypeptide comprised in the intracellular domain.Other portions of SEQ ID NO: 230 are shown in Table 20.

TABLE 20 nucleotide Sequence positions of SEQ number of Portions ID NO:230 nucleotides Kappa sp 210 . . . 269 60 LTR 1980 . . . 2449 470 M13fwd 3148 . . . 3164 17 AmpR 3639 . . . 3743 105 promoter AmpR 3744 . . .4604 861 ori 4775 . . . 5363 589 CAP binding 5651 . . . 5672 22 site lacpromoter 5687 . . . 5717 31 lac operator 5725 . . . 5741 17 M13 rev 5749. . . 5765 17 LTR 6174 . . . 6767 594 MMLV Psi 6830 . . . 7187 358 gag7252 . . . 15  417 (truncated)

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:231 encodes a BCMA-targeted CAR (designated as BCMA-targeted BBzCAR37) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:61, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:62, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD8 polypeptide having amino acids 137 to 207 of SEQ ID NO:226, and an intracellular domain comprising a CD3ξ polypeptidecomprising amino acid sequence of amino acids 52 to 163 of SEQ ID NO:195, and a co-stimulatory signaling region comprising a 4-1BBpolypeptide having the amino acids 214-255 of SEQ ID NO: 197. Nucleotidesequences 270-1028 of SEQ ID NO: 231 encodes the human scFv. Nucleotidesequences 1038-1250 of SEQ ID NO: 231 encodes the CD8 polypeptidecomprised in the transmembrane domain. Nucleotide sequences 1251-1376 ofSEQ ID NO: 231 encodes the 4-1BB polypeptide comprised in theintracellular domain. Nucleotide sequences 1377-1715 of SEQ ID NO: 231encodes the CD3zeta polypeptide comprised in the intracellular domain.Other portions of SEQ ID NO: 231 are shown in Table 21.

TABLE 21 nucleotide Sequence positions of SEQ number of Portions ID NO:231 nucleotides Kappa sp 210 . . . 269 60 LTR 1995 . . . 2464 470 M13fwd 3163 . . . 3179 17 AmpR 3654 . . . 3758 105 promoter AmpR 3759 . . .4619 861 ori 4790 . . . 5378 589 CAP binding 5666 . . . 5687 22 site lacpromoter 5702 . . . 5732 31 lac operator 5740 . . . 5756 17 M13 rev 5764. . . 5780 17 LTR 6189 . . . 6782 594 MMLV Psi 6845 . . . 7202 358 gag7267 . . . 15  417 (truncated)

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:232 encodes a BCMA-targeted CAR (designated as BCMA-targeted BBzCAR40) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:57, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:58, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD8 polypeptide having amino acids 137 to 207 of SEQ ID NO:226, and an intracellular domain comprising a CD3ξ polypeptidecomprising amino acid sequence of amino acids 52 to 163 of SEQ ID NO:195, and a co-stimulatory signaling region comprising a 4-1BBpolypeptide having the amino acids 214-255 of SEQ ID NO: 197. Nucleotidesequences 270-1024 of SEQ ID NO: 232 encodes the human scFv. Nucleotidesequences 1032-1244 of SEQ ID NO: 232 encodes the CD8 polypeptidecomprised in the transmembrane domain. Nucleotide sequences 1245-1370 ofSEQ ID NO: 232 encodes the 4-1BB polypeptide comprised in theintracellular domain. Nucleotide sequences 1371-1709 of SEQ ID NO: 232encodes the CD3zeta polypeptide comprised in the intracellular domain.Other portions of SEQ ID NO: 232 are shown in Table 22.

TABLE 22 nucleotide Sequence positions of SEQ number of Portions ID NO:232 nucleotides Kappa sp 210 . . . 269 60 LTR 1989 . . . 2458 470 M13fwd 3157 . . . 3173 17 AmpR 3648 . . . 3752 105 promoter AmpR 3753 . . .4613 861 ori 4784 . . . 5372 589 CAP binding 5660 . . . 5681 22 site lacpromoter 5696 . . . 5726 31 lac operator 5734 . . . 5750 17 M13 rev 5758. . . 5774 17 LTR 6183 . . . 6776 594 MMLV Psi 6839 . . . 7196 358 gag7261 . . . 15  417 (truncated)

The isolated nucleic acid molecule having the nucleotide sequence of SEQID NO:233 encodes a BCMA-targeted CAR (designated as BCMA-targeted BBzCAR54) comprising a human scFv that comprises a heavy chain variableregion comprising amino acids having the sequence set forth in SEQ IDNO:53, a light chain variable region comprising amino acids having thesequence set forth in SEQ ID NO:54, and a linker having an amino acidsequence of SEQ ID NO:69 positioned between the heavy chain variableregion and the light chain variable region, a transmembrane domaincomprising a CD8 polypeptide having amino acids 137 to 207 of SEQ ID NO:226, and an intracellular domain comprising a CD3ξ polypeptidecomprising amino acid sequence of amino acids 52 to 163 of SEQ ID NO:195, and a co-stimulatory signaling region comprising a 4-1BBpolypeptide having the amino acids 214-255 of SEQ ID NO: 197. Nucleotidesequences 270-1003 of SEQ ID NO: 233 encodes the human scFv. Nucleotidesequences 1011-1223 of SEQ ID NO: 233 encodes the CD8 polypeptidecomprised in the transmembrane domain. Nucleotide sequences 1224-1349 ofSEQ ID NO: 233 encodes the 4-1BB polypeptide comprised in theintracellular domain. Nucleotide sequences 1350-1688 of SEQ ID NO: 233encodes the CD3zeta polypeptide comprised in the intracellular domain.Other portions of SEQ ID NO: 233 are shown in Table 23.

TABLE 23 nucleotide Sequence positions of SEQ number of Portions ID NO:233 nucleotides Kappa sp 210 . . . 269 60 LTR 1968 . . . 2437 470 M13fwd 3136 . . . 3152 17 AmpR 3627 . . . 3731 105 promoter AmpR 3732 . . .4592 861 ori 4763 . . . 5351 589 CAP binding 5639 . . . 5660 22 site lacpromoter 5675 . . . 5705 31 lac operator 5713 . . . 5729 17 M13 rev 5737. . . 5753 17 LTR 6162 . . . 6755 594 MMLV Psi 6818 . . . 7175 358 gag7240 . . . 15  417 (truncated)

In certain embodiments, the isolated nucleic acid molecule encodes afunctional portion of a presently disclosed BCMA-targeted CAR. As usedherein, the term “functional portion” refers to any portion, part orfragment of a presently disclosed BCMA-targeted CAR, which portion, partor fragment retains the biological activity of the BCMA-targeted CAR(the parent CAR). For example, functional portions encompass theportions, parts or fragments of a presently disclosed BCMA-targeted CARthat retains the ability to recognize a target cell, to treat a disease,e.g., multiple myeloma, to a similar, same, or even a higher extent asthe parent CAR. In certain embodiments, an isolated nucleic acidmolecule encoding a functional portion of a presently disclosedBCMA-targeted CAR can encode a protein comprising, e.g., about 10%, 20%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,and 95%, or more of the parent CAR.

One Phase I clinical trial (NCT02215967) operated by National CancerInstitute (NCI) used anti-BCMA CAR-transduced T cells for treatingmultiple myeloma.^(33,34) The anti-BCMA CAR applied in the NCI'sclinical trial comprises a murine scFv binding to human BCMA. Using amouse antibody or a mouse scFv for treating humans can lead toanti-mouse antibody (HAMA) response, which may be life-threatening.Unlike NCI clinical trial, in certain embodiments, the presentlydisclosed BCMA-targeted CAR comprises a human scFv, and thus, affords amuch decreased risk of immunogenicity, compared with CARs comprisingmurine antibodies (see Maus et al., Cancer Immunol Res (2003);1(1):26-31), which reports that the potential immunogenicity of CARsderived from murine antibodies may be a safety issue for mRNA CARs).

III. Immunoresponsive Cells

The presently disclosed subject matter provides immunoresponsive cellsexpressing a CAR that comprises an extracellular antigen-binding domain,a transmembrane domain and an intracellular domain, where theextracellular antigen-binding domain specifically binds to BCMA (e.g.,human BCMA) as described above. The immunoresponsive cells can betransduced with a presently disclosed CAR such that the cells expressthe CAR. The presently disclosed subject matter also provides methods ofusing such cells for the treatment of a tumor, e.g., multiple myeloma(MM). The immunoresponsive cells of the presently disclosed subjectmatter can be cells of the lymphoid lineage. The lymphoid lineage,comprising B, T and natural killer (NK) cells, provides for theproduction of antibodies, regulation of the cellular immune system,detection of foreign agents in the blood, detection of cells foreign tothe host, and the like. Non-limiting examples of immunoresponsive cellsof the lymphoid lineage include T cells, Natural Killer (NK) cells,cytotoxic T lymphocytes (CTLs), regulatory T cells, embryonic stemcells, and pluripotent stem cells (e.g., those from which lymphoid cellsmay be differentiated). T cells can be lymphocytes that mature in thethymus and are chiefly responsible for cell-mediated immunity. T cellsare involved in the adaptive immune system. The T cells of the presentlydisclosed subject matter can be any type of T cells, including, but notlimited to, T helper cells, cytotoxic T cells, memory T cells (includingcentral memory T cells, stem-cell-like memory T cells (or stem-likememory T cells), and two types of effector memory T cells: e.g., TEMcells and TEMRA cells), Regulatory T cells (also known as suppressor Tcells), Natural killer T cells, Mucosal associated invariant T cells,and γδ T cells. In certain embodiments, the CAR-expressing T cellsexpress Foxp3 to achieve and maintain a T regulatory phenotype. Naturalkiller (NK) cells can be lymphocytes that are part of cell-mediatedimmunity and act during the innate immune response. NK cells do notrequire prior activation in order to perform their cytotoxic effect ontarget cells. Cytotoxic T cells (CTL or killer T cells) are a subset ofT lymphocytes capable of inducing the death of infected somatic or tumorcells.

The immunoresponsive cells of the presently disclosed subject matter canexpress an extracellular antigen-binding domain (e.g., a human scFV, aFab that is optionally crosslinked, or a F(ab)₂) that specifically bindsto BCMA (e.g., human BCMA), for the treatment of multiple myeloma. Suchimmunoresponsive cells can be administered to a subject (e.g., a humansubject) in need thereof for the treatment of multiple myeloma. Incertain embodiments, the immunoresponsive cell is a T cell. The T cellcan be a CD4⁺ T cell or a CD8⁺ T cell. In certain embodiments, the Tcell is a CD4⁺ T cell. In another embodiment, the T cell is a CD8⁺ Tcell.

A presently disclosed immunoresponsive cell can be further transducedwith at least one co-stimulatory ligand, such that the immunoresponsivecell co-expresses or is induced to co-express the BCMA-targeted CAR andthe at least one co-stimulatory ligand. The interaction between theBCMA-targeted CAR and at least one co-stimulatory ligand provides anon-antigen-specific signal important for full activation of animmunoresponsive cell (e.g., T cell). Co-stimulatory ligands include,but are not limited to, members of the tumor necrosis factor (TNF)superfamily, and immunoglobulin (Ig) superfamily ligands. TNF is acytokine involved in systemic inflammation and stimulates the acutephase reaction. Its primary role is in the regulation of immune cells.Members of TNF superfamily share a number of common features. Themajority of TNF superfamily members are synthesized as type IItransmembrane proteins (extracellular C-terminus) containing a shortcytoplasmic segment and a relatively long extracellular region. TNFsuperfamily members include, without limitation, nerve growth factor(NGF), CD40L (CD40L)/CD154, CD137L/4-1BBL, TNF-α, CD134L/OX40L/CD252,CD27L/CD70, Fas ligand (FasL), CD30L/CD153, tumor necrosis factor beta(TNFβ)/lymphotoxin-alpha (LTα), lymphotoxin-beta (LTβ), CD257/Bcell-activating factor (BAFF)/Blys/THANK/Tall-1, glucocorticoid-inducedTNF Receptor ligand (GITRL), and TNF-related apoptosis-inducing ligand(TRAIL), LIGHT (TNFSF14). The immunoglobulin (Ig) superfamily is a largegroup of cell surface and soluble proteins that are involved in therecognition, binding, or adhesion processes of cells. These proteinsshare structural features with immunoglobulins—they possess animmunoglobulin domain (fold). Immunoglobulin superfamily ligandsinclude, but are not limited to, CD80 and CD86, both ligands for CD28,PD-L1/(B7-H1) that ligands for PD-1. In certain embodiments, the atleast one co-stimulatory ligand is selected from the group consisting of4-1BBL, CD80, CD86, CD70, OX40L, CD48, TNFRSF14, PD-L1, and combinationsthereof. In certain embodiments, the immunoresponsive cell is transducedwith one co-stimulatory ligand that is 4-1BBL. In certain embodiments,the immunoresponsive cell is transduced with two co-stimulatory ligandsthat are 4-1BBL and CD80. CARs transduced with at least oneco-stimulatory ligand are described in U.S. Pat. No. 8,389,282, which isincorporated by reference in its entirety.

Furthermore, a presently disclosed immunoresponsive cell can be furthertransduced with at least one cytokine, such that the immunoresponsivecell secretes the at least one cytokine as well as expresses theBCMA-targeted CAR. In certain embodiments, the at least one cytokine isselected from the group consisting of IL-2, IL-3, IL-6, IL-7, IL-11,IL-12, IL-15, IL-17, and IL-21. In certain embodiments, the cytokine isIL-12.

The BCMA-specific or BCMA-targeted human lymphocytes that can be used inperipheral donor lymphocytes, e.g., those disclosed in Sadelain, M., etal. 2003 Nat Rev Cancer 3:35-45 (disclosing peripheral donor lymphocytesgenetically modified to express CARs), in Morgan, R. A., et al. 2006Science 314:126-129 (disclosing peripheral donor lymphocytes geneticallymodified to express a full-length tumor antigen-recognizing T cellreceptor complex comprising the α and β heterodimer), in Panelli, M. C.,et al. 2000 J Immunol 164:495-504; Panelli, M. C., et al. 2000 J Immunol164:4382-4392 (disclosing lymphocyte cultures derived from tumorinfiltrating lymphocytes (TILs) in tumor biopsies), and in Dupont, J.,et al. 2005 Cancer Res 65:5417-5427; Papanicolaou, G. A., et al. 2003Blood 102:2498-2505 (disclosing selectively in vitro-expandedantigen-specific peripheral blood leukocytes employing artificialantigen-presenting cells (AAPCs) or pulsed dendritic cells). Theimmunoresponsive cells (e.g., T cells) can be autologous, non-autologous(e.g., allogeneic), or derived in vitro from engineered progenitor orstem cells.

In certain embodiments, a presently disclosed immunoresponsive cell(e.g., T cell) expresses from about 1 to about 4, from about 2 to about4, from about 3 to about 4, from about 1 to about 2, from about 1 toabout 3, or from about 2 to about 3 vector copy numbers/cell of apresently disclosed BCMA-targeted CAR.

The unpurified source of CTLs may be any known in the art, such as thebone marrow, fetal, neonate or adult or other hematopoietic cell source,e.g., fetal liver, peripheral blood or umbilical cord blood. Varioustechniques can be employed to separate the cells. For instance, negativeselection methods can remove non-CTLs initially. Monoclonal antibodiesare particularly useful for identifying markers associated withparticular cell lineages and/or stages of differentiation for bothpositive and negative selections.

A large proportion of terminally differentiated cells can be initiallyremoved by a relatively crude separation. For example, magnetic beadseparations can be used initially to remove large numbers of irrelevantcells. Preferably, at least about 80%, usually at least 70% of the totalhematopoietic cells will be removed prior to cell isolation.

Procedures for separation include, but are not limited to, densitygradient centrifugation; resetting; coupling to particles that modifycell density; magnetic separation with antibody-coated magnetic beads;affinity chromatography; cytotoxic agents joined to or used inconjunction with a mAb, including, but not limited to, complement andcytotoxins; and panning with antibody attached to a solid matrix, e.g.plate, chip, elutriation or any other convenient technique.

Techniques for separation and analysis include, but are not limited to,flow cytometry, which can have varying degrees of sophistication, e.g.,a plurality of color channels, low angle and obtuse light scatteringdetecting channels, impedance channels.

The cells can be selected against dead cells, by employing dyesassociated with dead cells such as propidium iodide (PI). Preferably,the cells are collected in a medium comprising 2% fetal calf serum (FCS)or 0.2% bovine serum albumin (BSA) or any other suitable, preferablysterile, isotonic medium.

IV. Vectors

Genetic modification of immunoresponsive cells (e.g., T cells, CTLcells, NK cells) can be accomplished by transducing a substantiallyhomogeneous cell composition with a recombinant DNA or RNA construct.The vector can be a retroviral vector (e.g., gamma retroviral), which isemployed for the introduction of the DNA or RNA construct into the hostcell genome. For example, a polynucleotide encoding the BCMA-targetedCAR can be cloned into a retroviral vector and expression can be drivenfrom its endogenous promoter, from the retroviral long terminal repeat,or from an alternative internal promoter.

Non-viral vectors or RNA may be used as well. Random chromosomalintegration, or targeted integration (e.g., using a nuclease,transcription activator-like effector nucleases (TALENs), Zinc-fingernucleases (ZFNs), and/or clustered regularly interspaced shortpalindromic repeats (CRISPRs), or transgene expression (e.g., using anatural or chemically modified RNA) can be used.

For initial genetic modification of the cells to provide BCMA-targetedCAR expressing cells, a retroviral vector is generally employed fortransduction, however any other suitable viral vector or non-viraldelivery system can be used. For subsequent genetic modification of thecells to provide cells comprising an antigen presenting complexcomprising at least two co-stimulatory ligands, retroviral gene transfer(transduction) likewise proves effective. Combinations of retroviralvector and an appropriate packaging line are also suitable, where thecapsid proteins will be functional for infecting human cells. Variousamphotropic virus-producing cell lines are known, including, but notlimited to, PA12 (Miller, et al. (1985) Mol. Cell. Biol. 5:431-437);PA317 (Miller, et al. (1986) Mol. Cell. Biol. 6:2895-2902); and CRIP(Danos, et al. (1988) Proc. Natl. Acad. Sci. USA 85:6460-6464).Non-amphotropic particles are suitable too, e.g., particles pseudotypedwith VSVG, RD114 or GALV envelope and any other known in the art.

Possible methods of transduction also include direct co-culture of thecells with producer cells, e.g., by the method of Bregni, et al. (1992)Blood 80:1418-1422, or culturing with viral supernatant alone orconcentrated vector stocks with or without appropriate growth factorsand polycations, e.g., by the method of Xu, et al. (1994) Exp. Hemat.22:223-230; and Hughes, et al. (1992) J. Clin. Invest. 89:1817.

Transducing viral vectors can be used to express a co-stimulatory ligand(e.g., 4-1BBL and IL-12) in an immunoresponsive cell. Preferably, thechosen vector exhibits high efficiency of infection and stableintegration and expression (see, e.g., Cayouette et al., Human GeneTherapy 8:423-430, 1997; Kido et al., Current Eye Research 15:833-844,1996; Bloomer et al., Journal of Virology 71:6641-6649, 1997; Naldini etal., Science 272:263 267, 1996; and Miyoshi et al., Proc. Natl. Acad.Sci. U.S.A. 94:10319, 1997). Other viral vectors that can be usedinclude, for example, adenoviral, lentiviral, and adeno-associated viralvectors, vaccinia virus, a bovine papilloma virus, or a herpes virus,such as Epstein-Barr Virus (also see, for example, the vectors ofMiller, Human Gene Therapy 15-14, 1990; Friedman, Science 244:1275-1281,1989; Eglitis et al., BioTechniques 6:608-614, 1988; Tolstoshev et al.,Current Opinion in Biotechnology 1:55-61, 1990; Sharp, The Lancet337:1277-1278, 1991; Cornetta et al., Nucleic Acid Research andMolecular Biology 36:311-322, 1987; Anderson, Science 226:401-409, 1984;Moen, Blood Cells 17:407-416, 1991; Miller et al., Biotechnology7:980-990, 1989; Le Gal La Salle et al., Science 259:988-990, 1993; andJohnson, Chest 107:77S-83S, 1995). Retroviral vectors are particularlywell developed and have been used in clinical settings (Rosenberg etal., N. Engl. J. Med 323:370, 1990; Anderson et al., U.S. Pat. No.5,399,346).

In certain non-limiting embodiments, the vector expressing a presentlydisclosed BCMA-targeted CAR is a retroviral vector, e.g., a 293galv9retroviral vector.

Non-viral approaches can also be employed for the expression of aprotein in cell. For example, a nucleic acid molecule can be introducedinto a cell by administering the nucleic acid in the presence oflipofection (Feigner et al., Proc. Nat'l. Acad. Sci. U.S.A. 84:7413,1987; Ono et al., Neuroscience Letters 17:259, 1990; Brigham et al., Am.J. Med. Sci. 298:278, 1989; Staubinger et al., Methods in Enzymology101:512, 1983), asialoorosomucoid-polylysine conjugation (Wu et al.,Journal of Biological Chemistry 263:14621, 1988; Wu et al., Journal ofBiological Chemistry 264:16985, 1989), or by micro-injection undersurgical conditions (Wolff et al., Science 247:1465, 1990). Othernon-viral means for gene transfer include transfection in vitro usingcalcium phosphate, DEAE dextran, electroporation, and protoplast fusion.Liposomes can also be potentially beneficial for delivery of DNA into acell. Transplantation of normal genes into the affected tissues of asubject can also be accomplished by transferring a normal nucleic acidinto a cultivatable cell type ex vivo (e.g., an autologous orheterologous primary cell or progeny thereof), after which the cell (orits descendants) are injected into a targeted tissue or are injectedsystemically. Recombinant receptors can also be derived or obtainedusing transposases or targeted nucleases (e.g. Zinc finger nucleases,meganucleases, or TALE nucleases). Transient expression may be obtainedby RNA electroporation.

cDNA expression for use in polynucleotide therapy methods can bedirected from any suitable promoter (e.g., the human cytomegalovirus(CMV), simian virus 40 (SV40), or metallothionein promoters), andregulated by any appropriate mammalian regulatory element or intron(e.g. the elongation factor 1 α enhancer/promoter/intron structure). Forexample, if desired, enhancers known to preferentially direct geneexpression in specific cell types can be used to direct the expressionof a nucleic acid. The enhancers used can include, without limitation,those that are characterized as tissue- or cell-specific enhancers.Alternatively, if a genomic clone is used as a therapeutic construct,regulation can be mediated by the cognate regulatory sequences or, ifdesired, by regulatory sequences derived from a heterologous source,including any of the promoters or regulatory elements described above.

The resulting cells can be grown under conditions similar to those forunmodified cells, whereby the modified cells can be expanded and usedfor a variety of purposes.

V. Polypeptides and Analogs and Polynucleotides

Also included in the presently disclosed subject matter areextracellular antigen-binding domains that specifically binds to a BCMA(e.g., human BCMA) (e.g., an scFv (e.g., a human scFv), a Fab, or a(Fab)₂), CD3ζ, CD8, CD28, etc. polypeptides or fragments thereof, andpolynucleotides encoding thereof that are modified in ways that enhancetheir anti-tumor activity when expressed in an immunoresponsive cell.The presently disclosed subject matter provides methods for optimizingan amino acid sequence or a nucleic acid sequence by producing analteration in the sequence. Such alterations may comprise certainmutations, deletions, insertions, or post-translational modifications.The presently disclosed subject matter further comprises analogs of anynaturally-occurring polypeptide of the presently disclosed subjectmatter. Analogs can differ from a naturally-occurring polypeptide of thepresently disclosed subject matter by amino acid sequence differences,by post-translational modifications, or by both. Analogs of thepresently disclosed subject matter can generally exhibit at least about85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%,about 96%, about 97%, about 98%, about 99% or more identity with all orpart of a naturally-occurring amino, acid sequence of the presentlydisclosed subject matter. The length of sequence comparison is at least5, 10, 15, 20, 25, 50, 75, 100 or more amino acid residues. Again, in anexemplary approach to determining the degree of identity, a BLASTprogram may be used, with a probability score between e⁻³ and e⁻¹⁰⁰indicating a closely related sequence. Modifications comprise in vivoand in vitro chemical derivatization of polypeptides, e.g., acetylation,carboxylation, phosphorylation, or glycosylation; such modifications mayoccur during polypeptide synthesis or processing or following treatmentwith isolated modifying enzymes. Analogs can also differ from thenaturally-occurring polypeptides of the presently disclosed subjectmatter by alterations in primary sequence. These include geneticvariants, both natural and induced (for example, resulting from randommutagenesis by irradiation or exposure to ethanemethylsulfate or bysite-specific mutagenesis as described in Sambrook, Fritsch andManiatis, Molecular Cloning: A Laboratory Manual (2d ed.), CSH Press,1989, or Ausubel et al., supra). Also included are cyclized peptides,molecules, and analogs which contain residues other than L-amina acids,e.g., D-amino acids or non-naturally occurring or synthetic amino acids,e.g., beta ((3) or gamma (γ) amino acids.

In addition to full-length polypeptides, the presently disclosed subjectmatter also provides fragments of any one of the polypeptides or peptidedomains of the presently disclosed subject matter. A fragment can be atleast 5, 10, 13, or 15 amino acids. In certain embodiments, a fragmentis at least 20 contiguous amino acids, at least 30 contiguous aminoacids, or at least 50 contiguous amino acids. In certain embodiments, afragment is at least 60 to 80, 100, 200, 300 or more contiguous aminoacids. Fragments of the presently disclosed subject matter can begenerated by methods known to those of ordinary skill in the art or mayresult from normal protein processing (e.g., removal of amino acids fromthe nascent polypeptide that are not required for biological activity orremoval of amino acids by alternative mRNA splicing or alternativeprotein processing events).

Non-protein analogs have a chemical structure designed to mimic thefunctional activity of a protein of the invention. Such analogs areadministered according to methods of the presently disclosed subjectmatter. Such analogs may exceed the physiological activity of theoriginal polypeptide. Methods of analog design are well known in theart, and synthesis of analogs can be carried out according to suchmethods by modifying the chemical structures such that the resultantanalogs increase the anti-neoplastic activity of the originalpolypeptide when expressed in an immunoresponsive cell. These chemicalmodifications include, but are not limited to, substituting alternativeR groups and varying the degree of saturation at specific carbon atomsof a reference polypeptide. The protein analogs can be relativelyresistant to in vivo degradation, resulting in a more prolongedtherapeutic effect upon administration. Assays for measuring functionalactivity include, but are not limited to, those described in theExamples below.

In accordance with the presently disclosed subject matter, thepolynucleotides encoding an extracellular antigen-binding domain thatspecifically binds to BCMA (e.g., human BCMA) (e.g., an scFv (e.g., ahuman scFv), a Fab, or a (Fab)₂), CD3ζ, CD8, CD28) can be modified bycodon optimization. Codon optimization can alter both naturallyoccurring and recombinant gene sequences to achieve the highest possiblelevels of productivity in any given expression system. Factors that areinvolved in different stages of protein expression include codonadaptability, mRNA structure, and various cis-elements in transcriptionand translation. Any suitable codon optimization methods or technologiesthat are known to ones skilled in the art can be used to modify thepolynucleotids of the presently disclosed subject matter, including, butnot limited to, OptimumGene™, Encor optimization, and Blue Heron.

VI. Administration

BCMA-targeted CARs and immunoresponsive cells expressing thereof of thepresently disclosed subject matter can be provided systemically ordirectly to a subject for treating or preventing a neoplasia. In certainembodiments, the BCMA-targeted CARs and immunoresponsive cellsexpressing thereof are directly injected into an organ of interest(e.g., an organ affected by a neoplasia). Alternatively or additionally,the BCMA-targeted CARs and immunoresponsive cells expressing thereof areprovided indirectly to the organ of interest, for example, byadministration into the circulatory system (e.g., the tumorvasculature). Expansion and differentiation agents can be provided priorto, during or after administration of cells and compositions to increaseproduction of T cells in vitro or in vivo.

BCMA-targeted CARs and immunoresponsive cells expressing thereof of thepresently disclosed subject matter can be administered in anyphysiologically acceptable vehicle, normally intravascularly, althoughthey may also be introduced into bone or other convenient site where thecells may find an appropriate site for regeneration and differentiation(e.g., thymus). Usually, at least 1×10⁵ cells can be administered,eventually reaching 1×10¹⁰ or more. A cell population comprisingimmunoresponsive cells expressing a BCMA-targeted CAR can comprise apurified population of cells. Those skilled in the art can readilydetermine the percentage of immunoresponsive cells in a cell populationusing various well-known methods, such as fluorescence activated cellsorting (FACS). The ranges of purity in cell populations comprisinggenetically modified immunoresponsive cells expressing a BCMA-specificCAR can be from about 50% to about 55%, from about 55% to about 60%,from about 65% to about 70%, from about 70% to about 75%, from about 75%to about 80%, from about 80% to about 85%; from about 85% to about 90%,from about 90% to about 95%, or from about 95 to about 100%. Dosages canbe readily adjusted by those skilled in the art (e.g., a decrease inpurity may require an increase in dosage). The immunoresponsive cellscan be introduced by injection, catheter, or the like. If desired,factors can also be included, including, but not limited to,interleukins, e.g. IL-2, IL-3, IL 6, IL-11, IL-7, IL-12, IL-15, IL-21,as well as the other interleukins, the colony stimulating factors, suchas G-, M- and GM-CSF, interferons, e.g., γ-interferon.

Compositions of the presently disclosed subject matter comprisepharmaceutical compositions comprising immunoresponsive cells expressinga BCMA-targeted CAR and a pharmaceutically acceptable carrier.Administration can be autologous or non-autologous. For example,immunoresponsive cells expressing a BCMA-targeted CAR and compositionscomprising thereof can be obtained from one subject, and administered tothe same subject or a different, compatible subject. Peripheral bloodderived T cells of the presently disclosed subject matter or theirprogeny (e.g., in vivo, ex vivo or in vitro derived) can be administeredvia localized injection, including catheter administration, systemicinjection, localized injection, intravenous injection, or parenteraladministration. When administering a pharmaceutical composition of thepresently disclosed subject matter (e.g., a pharmaceutical compositioncomprising immunoresponsive cells expressing a BCMA-targeted CAR), itcan be formulated in a unit dosage injectable form (solution,suspension, emulsion).

VII. Formulations

Immunoresponsive cells expressing a generally BCMA-targeted CAR andcompositions comprising thereof of the presently disclosed subjectmatter can be conveniently provided as sterile liquid preparations,e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions,or viscous compositions, which may be buffered to a selected pH. Liquidpreparations are normally easier to prepare than gels, other viscouscompositions, and solid compositions. Additionally, liquid compositionsare somewhat more convenient to administer, especially by injection.Viscous compositions, on the other hand, can be formulated within theappropriate viscosity range to provide longer contact periods withspecific tissues. Liquid or viscous compositions can comprise carriers,which can be a solvent or dispersing medium containing, for example,water, saline, phosphate buffered saline, polyol (for example, glycerol,propylene glycol, liquid polyethylene glycol, and the like) and suitablemixtures thereof.

Sterile injectable solutions can be prepared by incorporating thecompositions comprising immunoresponsive cells expressing a generallyBCMA-targeted CAR of the presently disclosed subject matter in therequired amount of the appropriate solvent with various amounts of theother ingredients, as desired. Such compositions may be in admixturewith a suitable carrier, diluent, or excipient such as sterile water,physiological saline, glucose, dextrose, or the like. The compositionscan also be lyophilized. The compositions can contain auxiliarysubstances such as wetting, dispersing, or emulsifying agents (e.g.,methylcellulose), pH buffering agents, gelling or viscosity enhancingadditives, preservatives, flavoring agents, colors, and the like,depending upon the route of administration and the preparation desired.Standard texts, such as “REMINGTON'S PHARMACEUTICAL SCIENCE”, 17thedition, 1985, incorporated herein by reference, may be consulted toprepare suitable preparations, without undue experimentation.

Various additives which enhance the stability and sterility of thecompositions, including antimicrobial preservatives, antioxidants,chelating agents, and buffers, can be added. Prevention of the action ofmicroorganisms can be ensured by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, sorbic acid, andthe like. Prolonged absorption of the injectable pharmaceutical form canbe brought about by the use of agents delaying absorption, for example,alum inurn monostearate and gelatin. According to the present invention,however, any vehicle, diluent, or additive used would have to becompatible with the immunoresponsive cells expressing a generallyBCMA-targeted CAR of the presently disclosed subject matter.

The compositions can be isotonic, i.e., they can have the same osmoticpressure as blood and lacrimal fluid. The desired isotonicity of thecompositions of the presently disclosed subject matter may beaccomplished using sodium chloride, or other pharmaceutically acceptableagents such as dextrose, boric acid, sodium tartrate, propylene glycolor other inorganic or organic solutes. Sodium chloride is preferredparticularly for buffers containing sodium ions.

Viscosity of the compositions, if desired, can be maintained at theselected level using a pharmaceutically acceptable thickening agent.Methylcellulose can be used because it is readily and economicallyavailable and is easy to work with. Other suitable thickening agentsinclude, for example, xanthan gum, carboxymethyl cellulose,hydroxypropyl cellulose, carbomer, and the like. The concentration ofthe thickener can depend upon the agent selected. The important point isto use an amount that will achieve the selected viscosity. Obviously,the choice of suitable carriers and other additives will depend on theexact route of administration and the nature of the particular dosageform, e.g., liquid dosage form (e.g., whether the composition is to beformulated into a solution, a suspension, gel or another liquid form,such as a time release form or liquid-filled form).

Those skilled in the art will recognize that the components of thecompositions should be selected to be chemically inert and will notaffect the viability or efficacy of the immunoresponsive cells asdescribe in the presently disclosed subject matter. This will present noproblem to those skilled in chemical and pharmaceutical principles, orproblems can be readily avoided by reference to standard texts or bysimple experiments (not involving undue experimentation), from thisdisclosure and the documents cited herein.

One consideration concerning the therapeutic use of the immunoresponsivecells of the presently disclosed subject matter is the quantity of cellsnecessary to achieve an optimal effect. The quantity of cells to beadministered will vary for the subject being treated. In certainembodiments, from about 10⁴ to about 10¹⁰, from about 10⁵ to about 10⁹,or from about 10⁶ to about 10⁸ immunoresponsive cells of the presentlydisclosed subject matter are administered to a subject. More effectivecells may be administered in even smaller numbers. In certainembodiments, at least about 1×10⁸, about 2×10⁸, about 3×10⁸, about4×10⁸, and about 5×10⁸ immunoresponsive cells of the presently disclosedsubject matter are administered to a human subject. The precisedetermination of what would be considered an effective dose may be basedon factors individual to each subject, including their size, age, sex,weight, and condition of the particular subject. Dosages can be readilyascertained by those skilled in the art from this disclosure and theknowledge in the art.

The skilled artisan can readily determine the amount of cells andoptional additives, vehicles, and/or carrier in compositions and to beadministered in methods of the presently disclosed subject matter.Typically, any additives (in addition to the active cell(s) and/oragent(s)) are present in an amount of from about 0.001% to about 50% byweight) solution in phosphate buffered saline, and the active ingredientis present in the order of micrograms to milligrams, such as from about0.0001 wt % to about 5 wt %, from about 0.0001 wt % to about 1 wt %,from about 0.0001 wt % to about 0.05 wt %, from about 0.001 wt % toabout 20 wt %, from about 0.01 wt % to about 10 wt %, or from about 0.05wt % to about 5 wt %. For any composition to be administered to ananimal or human, and for any particular method of administration,toxicity should be determined, such as by determining the lethal dose(LD) and LD50 in a suitable animal model e.g., rodent such as mouse;and, the dosage of the composition(s), concentration of componentstherein and timing of administering the composition(s), which elicit asuitable response. Such determinations do not require undueexperimentation from the knowledge of the skilled artisan, thisdisclosure and the documents cited herein. And, the time for sequentialadministrations can be ascertained without undue experimentation.

VIII. Methods of Treatment

Tumor Microenvironment.

Tumors have a microenvironment that is hostile to the host immuneresponse involving a series of mechanisms by malignant cells to protectthemselves from immune recognition and elimination. This “hostile tumormicroenvironment” comprises a variety of immune suppressive factorsincluding infiltrating regulatory CD4⁺ T cells (Tregs), myeloid derivedsuppressor cells (MDSCs), tumor associated macrophages (TAMs), immunesuppressive cytokines including IL-10 and TGF-β, and expression ofligands targeted to immune suppressive receptors expressed by activatedT cells (CTLA-4 and PD-1). These mechanisms of immune suppression play arole in the maintenance of tolerance and suppressing inappropriateimmune responses, however within the tumor microenvironment thesemechanisms prevent an effective anti-tumor immune response. Collectivelythese immune suppressive factors can induce either marked anergy orapoptosis of adoptively transferred CAR modified T cells upon encounterwith targeted tumor cells.

Challenges in Tumor Immunology.

Effective tumor immunity requires recognition of tumor antigens andunopposed tumor elimination by immune effector cells. Tumor antigensmust contain peptide epitopes that are presented by the tumor and can berecognized by specific cytotoxic T lymphocytes (CTLs). The primed CTLsmust expand to a sufficient number and migrate to tumor sites, whereinthey mature into effectors to perform their functions, which areenhanced by helper T cells and dampened by Tregs and inhibitorymacrophages.

Targeted T Cell Therapy with Engineered T Lymphocytes.

T cell engineering is a groundbreaking strategy to potentially resolvemany previously observed shortcomings of earlier immunotherapeuticapproaches. Within the past year, researchers have reported dramaticcomplete remissions in relapsed^(16,17,) chemorefractory leukemia andmetastatic melanoma¹⁸⁻²⁰, obtained with autologous peripheral blood Tcells targeted to a defined antigen (CD19 and NY-ESO-1, respectively).

Rationale for a Genetic Approach:

Cell engineering can be used to redirect T cells toward tumor antigensand to enhance T cell function. One impetus for genetic T cellmodification is the potential to enhance T cell survival and expansionand to offset T cell death, anergy, and immune suppression. The genetictargeting of T cells can also be refined to prevent undesireddestruction of normal tissues.

Chimeric Antigen Receptors (CARs):

Tumor-specific T cells can be generated by the transfer of genes thatencode CARs²¹⁻²⁶. Second-generation CARs comprise a tumorantigen-binding domain fused to an intracellular signaling domaincapable of activating T cells and a co-stimulatory domain designed toaugment T cell potency and persistence²⁷. CAR design can thereforereconcile antigen recognition with signal transduction, two functionsthat are physiologically borne by two separate complexes, the TCRheterodimer and the CD3 complex. The CAR's extracellular antigen-bindingdomain is usually derived from a murine monoclonal antibody (mAb) orfrom receptors or their ligands. Antigen recognition is therefore notMHC-restricted^(28,29) and is therefore applicable to any patientexpressing the target antigen, using the same CAR. Antigen binding bythe CARs triggers phosphorylation of immunoreceptor tyrosine-basedactivation motifs (ITAMs) in the intracellular domain, initiating asignaling cascade required for cytolysis induction, cytokine secretion,and proliferation. Because MHC restriction of antigen recognition isbypassed, the function of CAR-targeted T cells is not affected by HLAdownregulation or defects in the antigen-processing machinery.

T Cell Requirements for Expansion and Survival:

Proliferation of tumor-specific T cells is needed ex vivo and isarguably desirable in vivo. T cell proliferation must be accompanied byT cell survival to permit absolute T cell expansion and persistence. Toproliferate in response to antigen, T cells must receive two signals.One is provided by TCR recognition of antigenic peptide/MHC complexesdisplayed on the surface of antigen-presenting cells (APCs)²⁵. The otheris provided by a T cell co-stimulatory receptor, such as the CD28 or4-1BB receptors. Whereas the cytolytic activity of T cells does notrequire concomitant co-stimulation, there is a critical need for theprovision of co-stimulatory signals to sustain the antitumor functionsof adoptively transferred T cells, as previouslydemonstrated^(23,27,30-32).

Immune Monitoring:

Lymphocytes are multifunctional “drugs” that exhibit dynamicallyevolving effects after infusion. Upon antigen encounter, tumor-specificT cells activate and/or release a variety of proteins that can triggertumor killing, T cell proliferation, and recruitment or immunomodulationof other immune cells. Thus, measuring which proteins are secreted fromwhich cells, in what quantity, and at what time point yields profoundinsights into why a particular patient is or is not responding andprovides critical feedback for designing more-effective trials. Theseassay systems will permit direct and meaningful comparisons of clinicalapproaches and thus help design rational, next-generation therapeuticstrategies.

For treatment, the amount administered is an amount effective inproducing the desired effect. An effective amount can be provided in oneor a series of administrations. An effective amount can be provided in abolus or by continuous perfusion.

An “effective amount” (or, “therapeutically effective amount”) is anamount sufficient to affect a beneficial or desired clinical result upontreatment. An effective amount can be administered to a subject in oneor more doses. In terms of treatment, an effective amount is an amountthat is sufficient to palliate, ameliorate, stabilize, reverse or slowthe progression of the disease, or otherwise reduce the pathologicalconsequences of the disease. The effective amount is generallydetermined by the physician on a case-by-case basis and is within theskill of one in the art. Several factors are typically taken intoaccount when determining an appropriate dosage to achieve an effectiveamount. These factors include age, sex and weight of the subject, thecondition being treated, the severity of the condition and the form andeffective concentration of the immunoresponsive cells administered.

For adoptive immunotherapy using antigen-specific T cells, cell doses inthe range of about 10⁶ to about 10¹⁰ (e.g., about 10⁹) are typicallyinfused. Upon administration of the immunoresponsive cells into thesubject and subsequent differentiation, the immunoresponsive cells areinduced that are specifically directed against one specific antigen(e.g., BCMA). “Induction” of T cells can include inactivation ofantigen-specific T cells such as by deletion or anergy. Inactivation isparticularly useful to establish or reestablish tolerance such as inautoimmune disorders. The immunoresponsive cells of the presentlydisclosed subject matter can be administered by any methods known in theart, including, but not limited to, pleural administration, intravenousadministration, subcutaneous administration, intranodal administration,intratumoral administration, intrathecal administration, intrapleuraladministration, intraperitoneal administration, and directadministration to the thymus. In certain embodiments, theimmunoresponsive cells and the compositions comprising thereof areintravenously administered to the subject in need.

The presently disclosed subject matter provides various methods of usingthe immunoresponsive cells (e.g., T cells) expressing a BCMA-targetedCAR. For example, the presently disclosed subject matter providesmethods of reducing tumor burden in a subject. In one non-limitingexample, the method of reducing tumor burden comprises administering aneffective amount of the presently disclosed immunoresponsive cell to thesubject, thereby inducing tumor cell death in the subject. The presentlydisclosed immunoresponsive cell can reduce the number of tumor cells,reduce tumor size, and/or eradicate the tumor in the subject.Non-limiting examples of suitable tumor include multiple myeloma,Non-Hodgkin Lymphoma, Hodgkin Lymphoma, Chronic Lymphocytic Leukemia(CLL), glioblastoma, and Waldenstrom's Macroglobulinemia. In certainembodiments, the tumor is multiple myeloma.

The presently disclosed subject matter also provides methods ofincreasing or lengthening survival of a subject having a neoplasia. Inone non-limiting example, the method of increasing or lengtheningsurvival of a subject having neoplasia comprises administering aneffective amount of the presently disclosed immunoresponsive cell to thesubject, thereby increasing or lengthening survival of the subject. Themethod can reduce or eradicate tumor burden in the subject. Thepresently disclosed subject matter further provides methods for treatingor preventing a neoplasia in a subject, comprising administering thepresently disclosed immunoresponsive cell to the subject.

As used herein, the term “neoplasia” refers to a disease characterizedby the pathological proliferation of a cell or tissue and its subsequentmigration to or invasion of other tissues or organs. Neoplasia growth istypically uncontrolled and progressive, and occurs under conditions thatwould not elicit, or would cause cessation of, multiplication of normalcells. Neoplasias can affect a variety of cell types, tissues, ororgans, including but not limited to an organ selected from the groupconsisting of bladder, colon, bone, brain, breast, cartilage, glia,esophagus, fallopian tube, gallbladder, heart, intestines, kidney,liver, lung, lymph node, nervous tissue, ovaries, pleura, pancreas,prostate, skeletal muscle, skin, spinal cord, spleen, stomach, testes,thymus, thyroid, trachea, urogenital tract, ureter, urethra, uterus, andvagina, or a tissue or cell type thereof. Neoplasias include cancers,such as sarcomas, carcinomas, or plasmacytomas (malignant tumor of theplasma cells).

Cancers whose growth may be inhibited using the immunoresponsive cellsof the presently disclosed subject matter comprise cancers typicallyresponsive to immunotherapy. Non-limiting examples of cancers fortreatment include multiple myeloma, Non-Hodgkin Lymphoma, HodgkinLymphoma, Chronic Lymphocytic Leukemia (CLL), glioblastoma, andWaldenstrom's Macroglobulinemia. In certain embodiments, the cancer ismultiple myeloma.

Additionally, the presently disclosed subject matter provides methods ofincreasing immune-activating cytokine production in response to a cancercell in a subject. In one non-limiting example, the method comprisesadministering the presently disclosed immunoresponsive cell to thesubject. The immune-activating cytokine can be granulocyte macrophagecolony stimulating factor (GM-CSF), IFN-α, IFN-β, IFN-γ, TNF-α, IL-2,IL-3, IL-6, IL-11, IL-7, IL-12, IL-15, IL-21, interferon regulatoryfactor 7 (IRF7), and combinations thereof. In certain embodiments, theimmunoresponsive cells including a BCMA-specific CAR of the presentlydisclosed subject matter increase the production of GM-CSF, IFN-γ,and/or TNF-α.

Suitable human subjects for therapy typically comprise two treatmentgroups that can be distinguished by clinical criteria. Subjects with“advanced disease” or “high tumor burden” are those who bear aclinically measurable tumor (e.g., multiple myeloma). A clinicallymeasurable tumor is one that can be detected on the basis of tumor mass(e.g., by palpation, CAT scan, sonogram, mammogram or X-ray; positivebiochemical or histopathologic markers on their own are insufficient toidentify this population). A pharmaceutical composition embodied in thepresently disclosed subject matter is administered to these subjects toelicit an anti-tumor response, with the objective of palliating theircondition. Ideally, reduction in tumor mass occurs as a result, but anyclinical improvement constitutes a benefit. Clinical improvementcomprises decreased risk or rate of progression or reduction inpathological consequences of the tumor (e.g., multiple myeloma).

A second group of suitable subjects is known in the art as the “adjuvantgroup.” These are individuals who have had a history of neoplasia (e.g.,multiple myeloma), but have been responsive to another mode of therapy.The prior therapy can have included, but is not restricted to, surgicalresection, radiotherapy, and traditional chemotherapy. As a result,these individuals have no clinically measurable tumor. However, they aresuspected of being at risk for progression of the disease, either nearthe original tumor site, or by metastases. This group can be furthersubdivided into high-risk and low-risk individuals. The subdivision ismade on the basis of features observed before or after the initialtreatment. These features are known in the clinical arts, and aresuitably defined for each different neoplasia. Features typical ofhigh-risk subgroups are those in which the tumor (e.g., multiplemyeloma) has invaded neighboring tissues, or who show involvement oflymph nodes. Another group has a genetic predisposition to neoplasia(e.g., multiple myeloma) but has not yet evidenced clinical signs ofneoplasia (e.g., multiple myeloma). For instance, women testing positivefor a genetic mutation associated with breast cancer, but still ofchildbearing age, can wish to receive one or more of the antigen-bindingfragments described herein in treatment prophylactically to prevent theoccurrence of neoplasia until it is suitable to perform preventivesurgery.

The subjects can have an advanced form of disease (e.g., multiplemyeloma), in which case the treatment objective can include mitigationor reversal of disease progression, and/or amelioration of side effects.The subjects can have a history of the condition, for which they havealready been treated, in which case the therapeutic objective willtypically include a decrease or delay in the risk of recurrence.

Further modification can be introduced to the BCMA-targetedCAR-expressing immunoresponsive cells (e.g., T cells) to avert orminimize the risks of immunological complications (known as “malignantT-cell transformation”), e.g., graft versus-host disease (GvHD), or whenhealthy tissues express the same target antigens as the tumor cells,leading to outcomes similar to GvHD. A potential solution to thisproblem is engineering a suicide gene into the CAR-expressing T cells.Suitable suicide genes include, but are not limited to, Herpes simplexvirus thymidine kinase (hsv-tk), inducible Caspase 9 Suicide gene(iCasp-9), and a truncated human epidermal growth factor receptor(EGFRt) polypeptide. In certain embodiments, the suicide gene is anEGFRt polypeptide. The EGFRt polypeptide can enable T cell eliminationby administering anti-EGFR monoclonal antibody (e.g., cetuximab). EGFRtcan be covalently joined to the 3′ terminus of the intracellular domainof the BCMA-targeted CAR. The suicide gene can be included within thevector comprising nucleic acids encoding the presently disclosedBCMA-targeted CARs. In this way, administration of a prodrug designed toactivate the suicide gene (e.g., a prodrug (e.g., AP1903 that canactivates iCasp-9) during malignant T-cell transformation (e.g., GVHD)triggers apoptosis in the suicide gene-activated CAR-expressing T cells.

IX. Kits

The presently disclosed subject matter provides kits for the treatmentor prevention of a neoplasia (e.g., multiple myeloma). In certainembodiments, the kit comprises a therapeutic or prophylactic compositioncontaining an effective amount of an immunoresponsive cell comprising aBCMA-targeted CAR in unit dosage form. In particular embodiments, thecells further expresses at least one co-stimulatory ligand. In certainembodiments, the kit comprises a sterile container which contains atherapeutic or prophylactic vaccine; such containers can be boxes,ampules, bottles, vials, tubes, bags, pouches, blister-packs, or othersuitable container forms known in the art. Such containers can be madeof plastic, glass, laminated paper, metal foil, or other materialssuitable for holding medicaments.

If desired, the immunoresponsive cell is provided together withinstructions for administering the cell to a subject having or at riskof developing a neoplasia (e.g., multiple myeloma). The instructionswill generally include information about the use of the composition forthe treatment or prevention of a neoplasia (e.g., multiple myeloma). Inother embodiments, the instructions include at least one of thefollowing: description of the therapeutic agent; dosage schedule andadministration for treatment or prevention of a neoplasia (e.g.,multiple myeloma) or symptoms thereof; precautions; warnings;indications; counter-indications; overdosage information; adversereactions; animal pharmacology; clinical studies; and/or references. Theinstructions may be printed directly on the container (when present), oras a label applied to the container, or as a separate sheet, pamphlet,card, or folder supplied in or with the container.

EXAMPLES

The practice of the present invention employs, unless otherwiseindicated, conventional techniques of molecular biology (includingrecombinant techniques), microbiology, cell biology, biochemistry andimmunology, which are well within the purview of the skilled artisan.Such techniques are explained fully in the literature, such as,“Molecular Cloning: A Laboratory Manual”, second edition (Sambrook,1989); “Oligonucleotide Synthesis” (Gait, 1984); “Animal Cell Culture”(Freshney, 1987); “Methods in Enzymology” “Handbook of ExperimentalImmunology” (Weir, 1996); “Gene Transfer Vectors for Mammalian Cells”(Miller and Calos, 1987); “Current Protocols in Molecular Biology”(Ausubel, 1987); “PCR: The Polymerase Chain Reaction”, (Mullis, 1994);“Current Protocols in Immunology” (Coligan, 1991). These techniques areapplicable to the production of the polynucleotides and polypeptides ofthe invention, and, as such, may be considered in making and practicingthe invention. Particularly useful techniques for particular embodimentswill be discussed in the sections that follow.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the assay, screening, and therapeutic methods of theinvention, and are not intended to limit the scope of what the inventorsregard as their invention.

Example 1BCMA Expression in Various Tissues

The Expression of human BCMA was evaluated in various malignant andnormal tissues by investigating gene expression profiles in databasessuch as the cancer cell line encyclopedia and BioGPS. As shown in FIGS.2A-2D, human BCMA was highly expressed in lymphoma and multiple myeloma,but not in other malignant tissues. Normal expression appeared limitedto B-cells and plasma cells. Potential BCMA targeted CAR T celleradication of these normal cell types may not have significant adverseeffects based on inventors' patient experience with CD19 targeted CAR Tcells. Any lack of physiologic antibody production can be addressed withintravenous immunoglobulin treatment.

Example 2—Construct of BCMA-Specific 28z CARs

Multiple unique fully human scFv's to BCMA were generated, and CARsbased on these scFv's were generated. Multiple scFv's were identified byscreening a fully human scFv phage library (>6×10¹⁰ scFv's) with BCMA-Fcfusion protein and then 3T3 cells expressing human BCMA. Aftersequencing, 57 unique and BCMA-Fc positive clones were found out of 79sequenced positive clones; the unique clone rate was 72%. FACS analysisof phage antibody clones against BCMA-3T3 and parental 3T3 cell linesresulted in confirming 25 unique positive clones.

ET140-153 scFv (or “ET140-3 scFv”), ET140-174 scFv (or “ET140-24 scFv”),ET140-187 scFv (or “ET140-37 scFv”), ET140-190 scFv (or “ET140-40scFv”), and ET140-204 scFv (or “ET140-54 scFv”) were used to generateBCMA-targeted 28z CARs 3, 24, 37, 40, and 54, respectively. TheseBCMA-targeted 28z CARs have similar structure, e.g., each has atransmembrane domain comprising a CD28 polypeptide, and an intracellulardomain comprising a CD3ξ polypeptide and a co-stimulatory signalingregion that comprises a CD28 polypeptide, as shown in FIG. 1. Each ofthese BCMA-targeted CARs were cloned into a retroviral vector. Theseviral vectors were then transduced into HEK 293galv9 viral packagingcells in order to generate a stable packaging line for generation ofCAR⁺ T cells.

Human T cells (unselected (CD4 and CD8) human T cells from a healthydonor) were transduced with retrovirus in order to express each of theseBCMA-targeted CARs such that the T cells expressed these BCMA-targeted28z CARs. The cell surface expression of BCMA-targeted CARs on human Tcells was determined via binding A647 conjugated BCMA-Fc fusion protein.The cell surface expression of BCMA-targeted 28z CAR24 was assessed, andcell surface detection was valided by flow cytometry, as shown in FIG.3.

The cross-reacting activity of seventeen human scFv's between human BCMAand mouse BCMA was assessed. As shown in FIG. 4, certain scFv's, e.g.,ET140-153 scFv (or “ET140-3 scFv”) and ET140-192 scFv (or “ET140-42scFv”) cross-reacted with mouse BCMA, thus, this scFv can be used forsyngeneic mouse studies.

Example 3—Activity of BCMA-Specific CARs

The anti-tumor activity of the presently disclosed BCMA-specific 28zCARs was evaluated. The in vitro data showed that the BCMA-specific CARsspecifically killed BCMA presenting cells, including MM cell lines. Forexample, as shown in FIG. 5, the T cells expressing the BCMA-specific28z CAR24 killed 3T3 cells overexpressing BCMA (but not control 3T3soverexpressing an irrelevant antigen). As shown in FIG. 6, the T cellsexpressing the BCMA-specific 28z CARs 24, 40, and 54 killed human MMcell lines.

Example 4—Screening Data for Anti-BCMA Antibodies

ELISA Screening: FIG. 21 shows the representative results of proteinELISA screening against BCMA antigen using specific scFv phage antibodyclones (ET140-3, ET140-24, ET140-37, ET140-40 and ET140-54). ELISAplates were coated with human BCMA ECD-Fc fusion protein, control-Fcfusion protein, or PBS alone as blank control, respectively. Individualphage clones from enriched phage display panning pools against BCMAECD-Fc fusion protein were incubated in the coated plates. Binding ofthe phage clones was detected by HRP-conjugated anti-M13 antibodies anddeveloped using TMB substrate. The absorbance was read at 450 nm.

FACS Screening: FIGS. 22A-22D show a representative figure of a FACSanalysis of the BCMA-specific phage antibody clones ET140-3, ET140-24,ET140-37, ET140-40 and ET140-54. Phage clones were incubated with3T3-BCMA cell line, then with anti-M13 mouse antibody. FinallyAPC-labeled anti-mouse IgG 2nd antibody was added to the reaction afterwashing again. The binding was measured by FACS and expressed as meanfluorescence intensity (MFI). Cells incubated with 2nd antibody alone,M13 K07 helper phage and cells only were used as negative controls.

Example 5—Construct of BCMA-Specific BBz CARs

Multiple unique fully human scFv's to BCMA were generated as describedin Example 2. ET140-153 scFv (or “ET140-3 scFv”), ET140-174 scFv (or“ET140-24 scFv”), ET140-187 scFv (or “ET140-37 scFv”), ET140-190 scFv(or “ET140-40 scFv”), and ET140-204 scFv (or “ET140-54 scFv”) were usedto generate BCMA-targeted BBz CARs 3, 24, 37, 40, and 54, respectively.Each of these BCMA-targeted BBz CARs has a transmembrane domaincomprising a CD8 polypeptide, and an intracellular domain comprising aCD3ξ polypeptide and a co-stimulatory signaling region that comprises a4-1BB polypeptide, as shown in FIG. 7. Each of these BCMA-targeted CARswere cloned into an SFG retroviral vector, as an example the 4-1BBcontaining CAR vectors are shown in FIGS. 8-12.

Example 6—Activity of BCMA-Targeted CAR T Cells

As shown in FIG. 13, BCMA-specific 28z CAR24 lysed human MM cell linesL363, NCL-H929, and U266, compared to irrelevantly targeted 4h11-28zMUC16 targeted CAR T cells. The cytotoxicity exhibited by observedBCMA-specific 28z CAR24 was specific to BCMA, as it did not lyse BCMAnegative CD19 positive Raji Burkett lymphoma cell line, as shown in FIG.13.

Example 7—Induction of Cytokine Secretion by BCMA-Targeted CAR T Cells

Co-culture of BCMA targeted 28z CAR24 T cells specifically with MM cellline induced cytokine secretion profile consistent with T cellactivation. FIG. 14 shows the IL-2 secretion after 24h co-culture of CART cells with human tumor cell lines (E:T ratio 1:1). Thelymphoplasmacytic lymphoma (CD19⁺) with CD19 targeted CAR T cells(positive control) and the MM cell line with the BCMA targeted 28z CAR24T cells displayed increased cytokine production. IFNg, IL-6, TNFa,sCD40L, GM-CSF all had similar secretion profiles (data not shown).

Example 8—Anti-Tumor Activity of BCMA-Targeted CAR T Cells

BCMA targeted 28z CAR54 T cells mediated an anti-myeloma immuneresponse. 1×10⁷ U266 human myeloma cell line cells were injected IV intoNSG mice on day 0. On day 4 1×10⁶ BCMA targeted or CD19 targeted secondgeneration CAR T cells were injected IV. Imaging on day 11 (day 7 s/pCAR T cell injection) shows that, unlike irrelevant (CD19) targeted CART cells; BCMA targeted 28z CAR54 T cells can mediate an anti-tumorresponse. See FIG. 15.

Example 9—Activity of BCMA-Targeted CAR T Cells

The ability of BCMA targeted CAR T cells to specifically lyse humanmyeloma cell line (HMCL) was tested. CD19 targeted CAR T cells or BCMAtargeted 28z CAR24 T cells were incubated with GFP expressing tumor celllines SET2 (Acute myeloid leukemia (AML), CD19⁻ BCMA⁻); BCWM1(Lymphoplasmacytic Lymphoma (LPL), CD19⁻BCMA⁻); L363 (Multiple Myeloma(MM), CD19⁻BCMA⁺). At time 0, the percent of GFP⁺ tumor line is shown inFIG. 16A. At 36h the positive control CD19 targeted CAR T cells havespecifically killed the GFP⁺LPL line, and similarly the BCMA targeted28z CAR24 T cells have specifically killed the GFP⁺ MM line. See FIG.16B.

Example 10Epitope Mapping of Anti-BCMA Antibodies

BCMA peptides were ordered based on the ECD sequence with N-terminalBiotin+SGSG linker+15 amino acids with 1 amino acid space. The peptidelibrary is shown in Table 24.

TABLE 24 ET140-pl SGSGLQMAGQCSQNEYFDS [SEQ ID NO: 236] ET140-p2SGSGQMAGQCSQNEYFDSL [SEQ ID NO: 237] ET140-p3SGSGMAGQCSQNEYFDSLL [SEQ ID NO: 238] ET140-p4SGSGAGQCSQNEYFDSLLH [SEQ ID NO: 239] ET140-p5SGSGGQCSQNEYFDSLLHA [SEQ ID NO: 240] ET140-p6SGSGQCSQNEYFDSLLHAC [SEQ ID NO: 241] ET140-p7SGSGCSQNEYFDSLLHACI [SEQ ID NO: 242] ET140-p8SGSGSQNEYFDSLLHACIP [SEQ ID NO: 243] ET140-p9SGSGQNEYFDSLLHACIPC [SEQ ID NO: 244] ET140-p10 SGSGNEYFDSLLHACIPCQ [SEQ ID NO: 245] ET140-pll SGSGEYFDSLLHACIPCQL [SEQ ID NO: 246] ET140-p12 SGSGYFDSLLHACIPCQLR [SEQ ID NO: 247] ET140-p13 SGSGFDSLLHACIPCQLRC [SEQ ID NO: 248] ET140-p14 SGSGDSLLHACIPCQLRCS [SEQ ID NO: 249] ET140-p15 SGSGSLLHACIPCQLRCSS [SEQ ID NO: 250] ET140-p16 SGSGLLHACIPCQLRCSSN [SEQ ID NO: 251] ET140-p17 SGSGLHACIPCQLRCSSNT [SEQ ID NO: 252] ET140-p18 SGSGHACIPCQLRCSSNTP [SEQ ID NO: 253] ET140-p19 SGSGACIPCQLRCSSNTPP [SEQ ID NO: 254] ET140-p20 SGSGCIPCQLRCSSNTPPL [SEQ ID NO: 255] ET140-p21 SGSGIPCQLRCSSNTPPLT [SEQ ID NO: 256] ET140-p22 SGSGPCQLRCSSNTPPLTC [SEQ ID NO: 257] ET140-p23 SGSGCQLRCSSNTPPLTCQ [SEQ ID NO: 258] ET140-p24 SGSGQLRCSSNTPPLTCQR [SEQ ID NO: 259] ET140-p25 SGSGLRCSSNTPPLTCQRY [SEQ ID NO: 260] ET140-p26 SGSGRCSSNTPPLTCQRYC [SEQ ID NO: 261] ET140-p27 SGSGCSSNTPPLTCQRYCN [SEQ ID NO: 262] ET140-p28 SGSGSSNTPPLTCQRYCNA [SEQ ID NO: 263] ET140-p29 SGSGSNTPPLTCQRYCNAS [SEQ ID NO: 264] ET140-p30 SGSGNTPPLTCQRYCNASV [SEQ ID NO: 265] ET140-p31 SGSGTPPLTCQRYCNASVT [SEQ ID NO: 266] ET140-p32 SGSGPPLTCQRYCNASVTN [SEQ ID NO: 267] ET140-p33 SGSGPLTCQRYCNASVTNS [SEQ ID NO: 268] ET140-p34 SGSGLTCQRYCNASVTNSV [SEQ ID NO: 269] ET140-p35 SGSGTCQRYCNASVTNSVK [SEQ ID NO: 270] ET140-p36 SGSGCQRYCNASVTNSVKG [SEQ ID NO: 271] ET140-p37 SGSGQRYCNASVTNSVKGT [SEQ ID NO: 272] ET140-p38 SGSGRYCNASVTNSVKGTN [SEQ ID NO: 273] ET140-p39 SGSGYCNASVTNSVKGTNA [SEQ ID NO: 274]

The peptides were coated onto Streptavidin plates at 2 ug/mL in PBST(PBS+0.05% Tween-20). After washing and blocking with 3% BSA. Afterwashing, 1 ug/mL ET140-3, ET140-24, ET140-54 or ET901 mIgG1 was added tothe wells, respectively. “mIgG1” used in all Examples represents thatthe variable region is fully human and the Fc part is mouse IgG1. ThenHRP anti-mouse IgG detection antibody was added to each well. Finally,the color was developed using TMB substrate. A450 was recorded for dataanalysis. The results are shown in FIGS. 17-20. As shown in FIGS. 17 and20, ET140-3 bound to peptides 7-13 (i.e., amino acids 8-22, 9-23, 10-24,11-25, 12-26, 13-27, and 14-28) of SEQ ID NO:71. As shown in FIGS. 18and 19, no linear epitopes found for ET140-24 or ET140-54.

Summary:

3 ET140 antibodies (mIgG1) were tested together with isotype controlET901 mIgG1 for their binding epitope towards BCMA-ECD. A peptidelibrary consisting of 39 peptides (N-terminal biotin+SGSG linker+15amino acids, with 1 amino acid offset) was used for epitope mappingELISA. This allows to search for the linear binding epitope of BCMA-ECD.ET901 mIgG1 was used as background reference for each peptide. OnlyET140-3 can be identified for its epitope region: a region comprisingamino acids 14-22 of SEQ ID NO:71, e.g., amino acids 8-28 of SEQ ID NO:71.

ET140-24 and ET140-54 did not show any significant binding towardspeptide library. This indicated that these two antibodies may recognizeconformational epitope rather than linear epitope of BCMA.

Example 11—Anti-BCMA Antibodies Recombinant Antigen by Surface PlasmonResonance

Kinetics of interaction between ET140-153 mIgG1 (or “ET140-3 mIgG1”),ET140-174 mIgG1 (or “ET140-24 mIgG1”), ET140-204 mIgG1 (or “ET140-54mIgG1”) and BCMA recombinant antigen was measured using a BIAcore ×100instrument. In brief, 50 μg/mL of modified streptavidin was immobilizedonto a Sensor Chip CAP by flowing the Biotin CAPture Reagent through theflow cells at 2 μL/min for 5 minutes. 10 ug/mL biotinylated BCMA-Fcprotein was loaded onto the flow cell at a rate of 30 μL/min for 3minutes. Following the standard protocol for kinetics, a series ofinjection of ESK1 was performed between 0.6 and 15 μg/mL, each stepconsisting of a 3 minute injection at 30 μL/min and 3 minutedisassociation. Afterwards, the surface was regenerated for 2 minuteswith a solution consisting of 75% v/v of 8M guanidine-HCl and 25% v/v 1MNaOH. Kinetic constants were derived by the global fitting (1:1 Langmuirbinding model) using BIAcore X100 Evaluation Software (Version 2.0.1).The binding affinity data are shown in Table 25.

TABLE 25 Protein KD ET140-24 mIgG1 KD: 4.8 nM (BiaCore) ET140-54 mIgG1KD: 8.1 nM (BiaCore) ET140-3 mIgG1 KD: 1.2 nM (BiaCore)

Although the foregoing presently disclosed subject matter has beendescribed in some detail by way of illustration and example for purposesof clarity of understanding, the descriptions and examples should not beconstrued as limiting the scope of the presently disclosed subjectmatter. The disclosures of all patent and scientific literature citedherein are expressly incorporated in their entirety by reference.

REFERENCES

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From the foregoing description, it will be apparent that variations andmodifications may be made to the invention described herein to adopt itto various usages and conditions. Such embodiments are also within thescope of the following claims.

All patents and publications and sequences referred to by accession orreference number mentioned in this specification are herein incorporatedby reference to the same extent as if each independent patent andpublication and sequence was specifically and individually indicated tobe incorporated by reference.

What is claimed:
 1. A nucleic acid molecule encoding a chimeric antigenreceptor (CAR) comprising an extracellular antigen-binding domain thatbinds to a B-cell maturation antigen (BCMA), a transmembrane domain, andan intracellular signaling domain, wherein the extracellularantigen-binding domain comprises: (a) a heavy chain variable regioncomprising a CDR1 comprising the amino acid sequence set forth in SEQ IDNO:89, a CDR2 comprising the amino acid sequence set forth in SEQ IDNO:90, and a CDR3 comprising the amino acid sequence set forth in SEQ IDNO:91; and a light chain variable region comprising a CDR1 comprisingthe amino acid sequence set forth in SEQ ID NO:92, a CDR2 comprising theamino acid sequence set forth in SEQ ID NO:93, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:94; (b) a heavy chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:95, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:96, and a CDR3 comprising the amino acid sequence setforth in SEQ ID NO:97; and a light chain variable region comprising aCDR1 comprising the amino acid sequence set forth in SEQ ID NO:98, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:99, and aCDR3 comprising the amino acid sequence set forth in SEQ ID NO:100; (c)a heavy chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:101, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:102, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:103; and a light chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:104, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:105, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:106; (d) a heavy chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:107, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:108, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:109;and a light chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:110, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:111, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:112; (e) a heavy chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:113, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:114, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:115; and a light chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:116, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:117, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:118;(f) a heavy chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:119, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:120, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:121; and a light chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:122, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:123, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:124; (g) a heavy chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:125, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:126, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:127;and a light chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:128, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:129, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:130; (h) a heavy chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:131, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:132, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:133; and a light chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:134, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:135, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:136;(i) a heavy chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:137, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:138, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:139; and a light chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:140, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:141, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:142; (j) a heavy chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:143, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:144, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:145;and a light chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:146, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:147, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:148; (k) a heavy chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:149, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:150, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:151; and a light chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:152, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:153, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:154;(l) a heavy chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:155, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:156, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:157; and a light chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:158, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:159, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:160; (m) a heavy chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:161, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:162, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:163;and a light chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:164, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:165, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:166; (n) a heavy chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:167, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:168, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:169; and a light chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:170, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:171, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:172;(o) a heavy chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:173, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:174, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:175; and a light chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:176, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:177, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:178; (p) a heavy chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:179, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:180, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:181;and a light chain variable region comprising a CDR1 comprising the aminoacid sequence set forth in SEQ ID NO:182, a CDR2 comprising the aminoacid sequence set forth in SEQ ID NO:183, and a CDR3 comprising theamino acid sequence set forth in SEQ ID NO:184; or (q) a heavy chainvariable region comprising a CDR1 comprising the amino acid sequence setforth in SEQ ID NO:185, a CDR2 comprising the amino acid sequence setforth in SEQ ID NO:186, and a CDR3 comprising the amino acid sequenceset forth in SEQ ID NO:187; and a light chain variable region comprisinga CDR1 comprising the amino acid sequence set forth in SEQ ID NO:188, aCDR2 comprising the amino acid sequence set forth in SEQ ID NO:189, anda CDR3 comprising the amino acid sequence set forth in SEQ ID NO:190. 2.A nucleic acid molecule encoding a chimeric antigen receptor (CAR)comprising an extracellular antigen-binding domain that binds to aB-cell maturation antigen (BCMA), a transmembrane domain, and anintracellular signaling domain, wherein the extracellularantigen-binding domain comprises: (a) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:1, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:2; (b) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:5, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:6; (c) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:9, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:10; (d) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:13, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:14; (e) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:17, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:18; (f) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:21, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:22; (g) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:25, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:26; (h) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:29, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:30; (i) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:33, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:34; (j) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:37, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:38; (k) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:41, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:42; (l) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:45, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:46; (m) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:49, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:50; (n) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:53, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:54; (o) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:57, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:58; (p) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:61, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:62; or (q) a heavy chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable regionsequence set forth in SEQ ID NO:65, and a light chain variable regioncomprising a CDR1, a CDR2, and a CDR3 of the light chain variable regionsequence set forth in SEQ ID NO:66.
 3. The nucleic acid molecule ofclaim 1, wherein: (a) the heavy chain variable region comprises an aminoacid sequence that has at least about 90% sequence identity to thesequence set forth in SEQ ID NO:1, and the light chain variable regioncomprises an amino acid sequence that has at least about 90% sequenceidentity to the sequence set forth in SEQ ID NO:2; (b) the heavy chainvariable region comprises an amino acid sequence that has at least about90% sequence identity to the sequence set forth in SEQ ID NO:5, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:6; (c) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:9, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:10; (d) the heavy chain variableregion comprises an amino acid sequence that has at least about 90%sequence identity to the sequence set forth in SEQ ID NO:13, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:14; (e) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:17, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:18; (f) the heavy chain variableregion comprises an amino acid sequence that has at least about 90%sequence identity to the sequence set forth in SEQ ID NO:21, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:22; (g) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:25, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:26; (h) the heavy chain variableregion comprises an amino acid sequence that has at least about 90%sequence identity to the sequence set forth in SEQ ID NO:29, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:30; (i) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:33, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:34; (j) the heavy chain variableregion comprises an amino acid sequence that has at least about 90%sequence identity to the sequence set forth in SEQ ID NO:37, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:38; (k) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:41, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:42; (l) the heavy chain variableregion comprises an amino acid sequence that has at least about 90%sequence identity to the sequence set forth in SEQ ID NO:45, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:46; (m) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:49, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:50; (n) the heavy chain variableregion comprises an amino acid sequence that has at least about 90%sequence identity to the sequence set forth in SEQ ID NO:53, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:54; (o) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:57, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:58; (p) the heavy chain variableregion comprises an amino acid sequence that has at least about 90%sequence identity to the sequence set forth in SEQ ID NO:61, and thelight chain variable region comprises an amino acid sequence that has atleast about 90% sequence identity to the sequence set forth in SEQ IDNO:62; or (q) the heavy chain variable region comprises an amino acidsequence that has at least about 90% sequence identity to the sequenceset forth in SEQ ID NO:65, and the light chain variable region comprisesan amino acid sequence that has at least about 90% sequence identity tothe sequence set forth in SEQ ID NO:66.
 4. The nucleic acid molecule ofclaim 1, wherein: (a) the heavy chain variable region comprises theamino acid sequence set forth in SEQ ID NO:1, and the light chainvariable region comprises the amino acid sequence set forth in SEQ IDNO:2; (b) the heavy chain variable region comprises the amino acidsequence set forth in SEQ ID NO:5, and the light chain variable regioncomprises the amino acid sequence set forth in SEQ ID NO:6; (c) theheavy chain variable region comprises the amino acid sequence set forthin SEQ ID NO:9, and the light chain variable region comprises the aminoacid sequence set forth in SEQ ID NO:10; (d) the heavy chain variableregion comprises the amino acid sequence set forth in SEQ ID NO:13, andthe light chain variable region comprises the amino acid sequence setforth in SEQ ID NO:14; (e) the heavy chain variable region comprises theamino acid sequence set forth in SEQ ID NO:17, and the light chainvariable region comprises the amino acid sequence set forth in SEQ IDNO:18; (f) the heavy chain variable region comprises the amino acidsequence set forth in SEQ ID NO:21, and the light chain variable regioncomprises the amino acid sequence set forth in SEQ ID NO:22; (g) theheavy chain variable region comprises the amino acid sequence set forthin SEQ ID NO:25, and the light chain variable region comprises the aminoacid sequence set forth in SEQ ID NO:26; (h) the heavy chain variableregion comprises the amino acid sequence set forth in SEQ ID NO:29, andthe light chain variable region comprises the amino acid sequence setforth in SEQ ID NO:30; (i) the heavy chain variable region comprises theamino acid sequence set forth in SEQ ID NO:33, and the light chainvariable region comprises the amino acid sequence set forth in SEQ IDNO:34; (j) the heavy chain variable region comprises the amino acidsequence set forth in SEQ ID NO:37, and the light chain variable regioncomprises the amino acid sequence set forth in SEQ ID NO:38; (k) theheavy chain variable region comprises the amino acid sequence set forthin SEQ ID NO:41, and the light chain variable region comprises the aminoacid sequence set forth in SEQ ID NO:42; (l) the heavy chain variableregion comprises the amino acid sequence set forth in SEQ ID NO:45, andthe light chain variable region comprises the amino acid sequence setforth in SEQ ID NO:46; (m) the heavy chain variable region comprises theamino acid sequence set forth in SEQ ID NO:49, and the light chainvariable region comprises the amino acid sequence set forth in SEQ IDNO:50; (n) the heavy chain variable region comprises the amino acidsequence set forth in SEQ ID NO:53, and the light chain variable regioncomprises the amino acid sequence set forth in SEQ ID NO:54; (o) theheavy chain variable region comprises the amino acid sequence set forthin SEQ ID NO:57, and the light chain variable region comprises the aminoacid sequence set forth in SEQ ID NO:58; (p) the heavy chain variableregion comprises the amino acid sequence set forth in SEQ ID NO:61, andthe light chain variable region comprises the amino acid sequence setforth in SEQ ID NO:62; or (q) the heavy chain variable region comprisesthe amino acid sequence set forth in SEQ ID NO:65, and the light chainvariable region comprises the amino acid sequence set forth in SEQ IDNO:66.
 5. The nucleic acid molecule of claim 2, wherein the heavy chainvariable region comprises a CDR1, a CDR2, and a CDR3 sequence set forthin SEQ ID NO:119, SEQ ID NO:120, and SEQ ID NO:121, respectively; andthe light chain variable region comprises a CDR1, a CDR2, and a CDR3sequence set forth in SEQ ID NO:122, SEQ ID NO:123, and SEQ ID NO:124,respectively.
 6. The nucleic acid molecule of claim 2, wherein the heavychain variable region comprises a CDR1, a CDR2, and a CDR3 sequence setforth in SEQ ID NO:167, SEQ ID NO:168, and SEQ ID NO:169, respectively;and the light chain variable region comprises a CDR1, a CDR2, and a CDR3sequence set forth in SEQ ID NO:170, SEQ ID NO:171, and SEQ ID NO:172,respectively.
 7. The nucleic acid molecule of claim 2, wherein the heavychain variable region comprises a CDR1, a CDR2, and a CDR3 sequence setforth in SEQ ID NO:185, SEQ ID NO:186, and SEQ ID NO:187, respectively;and the light chain variable region comprises a CDR1, a CDR2, and a CDR3sequence set forth in SEQ ID NO:188, SEQ ID NO:189, and SEQ ID NO:190,respectively.
 8. The nucleic acid molecule of claim 1, wherein theextracellular antigen-binding domain comprises a single-chain variablefragment (scFv).
 9. The nucleic acid molecule of claim 2, wherein theextracellular antigen-binding domain comprises a single-chain variablefragment (scFv).
 10. The nucleic acid molecule of claim 4, wherein theextracellular antigen-binding domain comprises a single-chain variablefragment (scFv).
 11. The nucleic acid molecule of claim 8, wherein thescFv is a human scFv.
 12. The nucleic acid molecule of claim 1, whereinthe extracellular antigen-binding domain comprises the amino acidsequence set forth in SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ IDNO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ IDNO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ IDNO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88.
 13. The nucleic acidmolecule of claim 1, comprising the nucleotide sequence set forth in SEQID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:230, SEQ ID NO:231,SEQ ID NO:232, SEQ ID NO:233, SEQ ID NO:234, or SEQ ID NO:235.
 14. Thenucleic acid molecule of claim 1, wherein the BCMA comprises the aminoacid sequence set forth in SEQ ID NO:71.
 15. The nucleic acid moleculeof claim 1, wherein the extracellular antigen-binding domain binds to anon-linear epitope of BCMA.
 16. The nucleic acid molecule of claim 1,wherein the extracellular antigen-binding domain binds to BCMA with abinding affinity (K_(D)) of from about 1×10⁻⁹M to about 1×10⁻⁸M.
 17. Thenucleic acid molecule of claim 1, wherein the intracellular signalingdomain comprises a signaling domain of a CD3ζ polypeptide.
 18. Thenucleic acid molecule of claim 17, wherein the intracellular signalingdomain further comprises at least one co-stimulatory signaling region.19. The nucleic acid molecule of claim 18, wherein the at least oneco-stimulatory signaling region comprises an intracellular signalingregion of a CD28 polypeptide, an intracellular signaling region of a4-1BB polypeptide, an intracellular signaling region of an OX40polypeptide, an intracellular signaling region of an ICOS polypeptide,an intracellular signaling region of a DAP-10 polypeptide, or acombination thereof.
 20. The nucleic acid molecule of claim 18, whereinthe at least one co-stimulatory signaling region comprises anintracellular signaling region of a CD28 polypeptide or an intracellularsignaling region of a 4-1BB polypeptide.
 21. The nucleic acid moleculeof claim 17, wherein the transmembrane domain comprises a CD8polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide,a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide,a BTLA polypeptide, a synthetic peptide, or a combination thereof. 22.The nucleic acid molecule of claim 17, wherein the transmembrane domaincomprises a transmembrane domain of a CD28 polypeptide.
 23. The nucleicacid molecule of claim 19, wherein the transmembrane domain comprises aCD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOSpolypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide,or a combination thereof.
 24. The nucleic acid molecule of claim 1,wherein the transmembrane domain comprises a transmembrane domain of aCD28 polypeptide, and the intracellular signaling domain comprises asignaling domain of a CD3ζ polypeptide and at least one co-stimulatorysignaling region that comprises an intracellular signaling region of a4-1BB polypeptide.
 25. The nucleic acid molecule of claim 2, wherein thetransmembrane domain comprises a transmembrane domain of a CD28polypeptide, and the intracellular signaling domain comprises asignaling domain of a CD3ζ polypeptide and at least one co-stimulatorysignaling region that comprises an intracellular signaling region of a4-1BB polypeptide.
 26. A vector comprising the nucleic acid molecule ofclaim
 1. 27. A host cell comprising the nucleic acid of claim
 1. 28. Thehost cell of claim 27, wherein the host cell is a T cell.
 29. A methodfor producing a cell that binds to B-cell maturation antigen (BCMA),comprising introducing into the cell the nucleic acid molecule ofclaim
 1. 30. The method of claim 29, wherein the cell is a T cell.